Novel compounds as inhibitors of hepatitis C virus NS3 serine protease

ABSTRACT

The present invention discloses novel compounds which have HCV protease inhibitory activity as well as methods for preparing such compounds. In another embodiment, the invention discloses pharmaceutical compositions comprising such compounds as well as methods of using them to treat disorders associated with the HCV protease.

This application is a divisional of U.S. application Ser. No.11/065,531, filed Feb. 24, 2005, and claims the benefit of U.S.Provisional Application, Ser. No. 60/548,535 filed Feb. 27, 2004.

FIELD OF THE INVENTION

The present invention relates to novel hepatitis C virus (“HCV”)protease inhibitors, pharmaceutical compositions containing one or moresuch inhibitors, methods of preparing and using such inhibitors to treathepatitis C and related disorders.

BACKGROUND OF THE INVENTION

Hepatitis C virus (HCV) is a (+)-sense single-stranded RNA virus thathas been implicated as the major causative agent in non-A, non-Bhepatitis (NANBH), particularly in blood-associated NANBH (BB-NANBH)(see, International Patent Application Publication No. WO 89/04669 andEuropean Patent Application Publication No. EP 381 216). NANBH is to bedistinguished from other types of viral-induced liver disease, such ashepatitis A virus (HAV), hepatitis B virus (HBV), delta hepatitis virus(HDV), cytomegalovirus (CMV) and Epstein-Barr virus (EBV), as well asfrom other forms of liver disease such as alcoholism and primary biliarcirrhosis.

Recently, an HCV protease necessary for polypeptide processing and viralreplication has been identified, cloned and expressed. (See, e.g. U.S.Pat. No. 5,712,145). This approximately 3000 amino acid polyproteincontains, from the amino terminus to the carboxy terminus, anucleocapsid protein (C), envelope proteins (E1 and E2) and severalnon-structural proteins (NS1, 2, 3, 4a, 5a and 5b). NS3 is anapproximately 68 kda protein, encoded by approximately 1893 nucleotidesof the HCV genome, and has two distinct domains: (a) a serine proteasedomain consisting of approximately 200 of the N-terminal amino acids;and (b) an RNA-dependent ATPase domain at the C-terminus of the protein.The NS3 protease is considered a member of the chymotrypsin familybecause of similarities in protein sequence, overall three-dimensionalstructure and mechanism of catalysis. Other chymotrypsin-like enzymesare elastase, factor Xa, thrombin, trypsin, plasmin, urokinase, tPA andPSA. The HCV NS3 serine protease is responsible for proteolysis of thepolypeptide (polyprotein) at the NS3/NS4a, NS4a/NS4b, NS4b/NS5a andNS5a/NS5b junctions and is thus responsible for generating four viralproteins during viral replication. This has made the HCV NS3 serineprotease an attractive target for antiviral chemotherapy. The inventivecompounds can inhibit such protease. They also can modulate theprocessing of hepatitis C virus (HCV) polypeptide.

It has been determined that the NS4a protein, an approximately 6 kdapolypeptide, is a co-factor for the serine protease activity of NS3.Autocleavage of the NS3/NS4a junction by the NS3/NS4a serine proteaseoccurs intramolecularly (i.e., cis) while the other cleavage sites areprocessed intermolecularly (i.e., trans).

Analysis of the natural cleavage sites for HCV protease revealed thepresence of cysteine at P1 and serine at P1′ and that these residues arestrictly conserved in the NS4a/NS4b, NS4b/NS5a and NS5a/NS5b junctions.The NS3/NS4a junction contains a threonine at P1 and a serine at P1′.The Cys→Thr substitution at NS3/NS4a is postulated to account for therequirement of cis rather than trans processing at this junction. See,e.g., Pizzi et al. (1994) Proc. Natl. Acad. Sci. (USA) 91:888-892,Failla et al. (1996) Folding & Design 1:35-42. The NS3/NS4a cleavagesite is also more tolerant of mutagenesis than the other sites. See,e.g., Kollykhalov et al. (1994) J. Virol. 68:7525-7533. It has also beenfound that acidic residues in the region upstream of the cleavage siteare required for efficient cleavage. See, e.g., Komoda et al. (1994) J.Virol. 68:7351-7357.

Inhibitors of HCV protease that have been reported include antioxidants(see, International Patent Application Publication No. WO 98/14181),certain peptides and peptide analogs (see, International PatentApplication Publication No. WO 98/17679, Landro et al. (1997) Biochem.36:9340-9348, Ingallinella et al. (1998) Biochem. 37:8906-8914,Llinàs-Brunet et al. (1998) Bioorg. Med. Chem. Lett. 8:1713-1718),inhibitors based on the 70-amino acid polypeptide eglin c (Martin et al.(1998) Biochem. 37:11459-11468, inhibitors affinity selected from humanpancreatic secretory trypsin inhibitor (hPSTI-C3) and minibodyrepertoires (MBip) (Dimasi et al. (1997) J. Virol. 71:7461-7469),cV_(H)E2 (a “camelized” variable domain antibody fragment) (Martin etal. (1997) Protein Eng. 10:607-614), and α1-antichymotrypsin (ACT)(Elzouki et al.) (1997) J. Hepat. 27:42-28). A ribozyme designed toselectively destroy hepatitis C virus RNA has recently been disclosed(see, BioWorld Today 9(217): 4 (Nov. 10, 1998)).

Reference is also made to the PCT Publications, No. WO 98/17679,published Apr. 30, 1998 (Vertex Pharmaceuticals Incorporated); WO98/22496, published May 28, 1998 (F. Hoffmann-La Roche AG); and WO99/07734, published Feb. 18, 1999 (Boehringer Ingelheim Canada Ltd.).

HCV has been implicated in cirrhosis of the liver and in induction ofhepatocellular carcinoma. The prognosis for patients suffering from HCVinfection is currently poor. HCV infection is more difficult to treatthan other forms of hepatitis due to the lack of immunity or remissionassociated with HCV infection. Current data indicates a less than 50%survival rate at four years post cirrhosis diagnosis. Patients diagnosedwith localized resectable hepatocellular carcinoma have a five-yearsurvival rate of 10-30%, whereas those with localized unresectablehepatocellular carcinoma have a five-year survival rate of less than 1%.

Reference is made to WO 00/59929 (U.S. Pat. No. 6,608,027, Assignee:Boehringer Ingelheim (Canada) Ltd.; Published Oct. 12, 2000) whichdiscloses peptide derivatives of the formula:

Reference is made to A. Marchetti et al, Synlett, S1, 1000-1002 (1999)describing the synthesis of bicylic analogs of an inhibitor of HCV NS3protease. A compound disclosed therein has the formula:

Reference is also made to W. Han et al, Bioorganic & Medicinal Chem.Lett, (2000) 10, 711-713, which describes the preparation of certainα-ketoamides, α-ketoesters and α-diketones containing allyl and ethylfunctionalities.

Reference is also made to WO 00/09558 (Assignee: Boehringer IngelheimLimited; Published Feb. 24, 2000) which discloses peptide derivatives ofthe formula:

where the various elements are defined therein. An illustrative compoundof that series is:

Reference is also made to WO 00/09543 (Assignee: Boehringer IngelheimLimited; Published Feb. 24, 2000) which discloses peptide derivatives ofthe formula:

where the various elements are defined therein. An illustrative compoundof that series is:

Reference is also made to U.S. Pat. No. 6,608,027 (Boehringer Ingelheim,Canada) which discloses NS3 protease inhibitors of the type:

wherein the various moieties are defined therein.

Current therapies for hepatitis C include interferon-α (INF_(α)) andcombination therapy with ribavirin and interferon. See, e.g., Beremgueret al. (1998) Proc. Assoc. Am. Physicians 110(2):98-112. These therapiessuffer from a low sustained response rate and frequent side effects.See, e.g., Hoofnagle et al. (1997) N. Engl. J. Med. 336:347. Currently,no vaccine is available for HCV infection.

Reference is further made to WO 01/74768 (Assignee: VertexPharmaceuticals Inc) published Oct. 11, 2001, which discloses certaincompounds of the following general formula (R is defined therein) asNS3-serine protease inhibitors of Hepatitis C virus:

A specific compound disclosed in the afore-mentioned WO 01/74768 has thefollowing formula:

PCT Publications WO 01/77113; WO 01/081325; WO 02/08198; WO 02/08256; WO02/08187; WO 02/08244; WO 02/48172; WO 02/08251; and pending U.S. patentapplication, Ser. No. 10/052,386, filed Jan. 18, 2002, disclose varioustypes of peptides and/or other compounds as NS-3 serine proteaseinhibitors of hepatitis C virus. The disclosures of those applicationsare incorporated herein by reference thereto.

There is a need for new treatments and therapies for HCV infection.There is a need for compounds useful in the treatment or prevention oramelioration of one or more symptoms of hepatitis C.

There is a need for methods of treatment or prevention or ameliorationof one or more symptoms of hepatitis C.

There is a need for methods for modulating the activity of serineproteases, particularly the HCV NS3/NS4a serine protease, using thecompounds provided herein.

There is a need for methods of modulating the processing of the HCVpolypeptide using the compounds provided herein.

SUMMARY OF THE INVENTION

In its many embodiments, the present invention provides a novel class ofinhibitors of the HCV protease, pharmaceutical compositions containingone or more of the compounds, methods of preparing pharmaceuticalformulations comprising one or more such compounds, and methods oftreatment or prevention of HCV or amelioration of one or more of thesymptoms of hepatitis C using one or more such compounds or one or moresuch formulations. Also provided are methods of modulating theinteraction of an HCV polypeptide with HCV protease. Among the compoundsprovided herein, compounds that inhibit HCV NS3/NS4a serine proteaseactivity are preferred. The present invention discloses compounds, orenantiomers, stereoisomers, rotamers, tautomers, diastereomers orracemates of said compounds, or a pharmaceutically acceptable salt,solvate or ester of said compounds, said compounds having the generalstructure shown in structural Formula 1:

wherein:

R¹ is H, OR⁸, NR⁹R¹⁰, or CHR⁹R¹⁰, wherein R⁸, R⁹ and R¹⁰ can be the sameor different, each being independently selected from the groupconsisting of H, alkyl-, alkenyl-, alkynyl-, aryl-, heteroalkyl-,heteroaryl-, cycloalkyl-, heterocyclyl-, arylalkyl-, andheteroarylalkyl;

A and M can be the same or different, each being independently selectedfrom R, OR, NHR, NRR′, SR, SO₂R, and halo; or A and M are connected toeach other such that the moiety:

shown above in Formula I forms either a three, four, six, seven oreight-membered cycloalkyl, a four to eight-membered heterocyclyl, a sixto ten-membered aryl, or a five to ten-membered heteroaryl;

E is C(H) or C(R);

L is C(H), C(R), CH₂C(R), or C(R)CH₂;

R, R′, R², and R³ can be the same or different, each being independentlyselected from the group consisting of H, alkyl-, alkenyl-, alkynyl-,cycloalkyl-, heteroalkyl-, heterocyclyl-, aryl-, heteroaryl-,(cycloalkyl)alkyl-, (heterocyclyl)alkyl-, aryl-alkyl-, andheteroaryl-alkyl-; or alternately R and R′ in NRR′ are connected to eachother such that NRR′ forms a four to eight-membered heterocyclyl;

and Y is selected from the following moieties:

wherein G is NH or O; and R¹⁵, R¹⁶, R¹⁷, R¹⁸, and R¹⁹ can be the same ordifferent, each being independently selected from the group consistingof H, alkyl, heteroalkyl, alkenyl, heteroalkenyl, alkynyl,heteroalkynyl, cycloalkyl, heterocyclyl, aryl, arylalkyl, heteroaryl,and heteroarylalkyl, or alternately, (i) either R¹⁵ and R¹⁶ areconnected to each other to form a four to eight-membered cyclicstructure, or R¹⁵ and R¹⁹ are connected to each other to form a four toeight-membered cyclic structure, and (ii) likewise, independently, R¹⁷and R¹⁸ are connected to each other to form a three to eight-memberedcycloalkyl or heterocyclyl;

wherein each of said alkyl, aryl, heteroaryl, cycloalkyl or heterocyclylcan be unsubstituted or optionally independently substituted with one ormore moieties selected from the group consisting of: hydroxy, alkoxy,aryloxy, thio, alkylthio, arylthio, amino, amido, alkylamino, arylamino,alkylsulfonyl, arylsulfonyl, sulfonamido, alkylsulfonamido,arylsulfonamido, alkyl, aryl, heteroaryl, keto, carboxy, carbalkoxy,carboxamido, alkoxycarbonylamino, alkoxycarbonyloxy, alkylureido,arylureido, halo, cyano, and nitro.

The above-noted statement “A and M are connected to each other such thatthe moiety:

shown above in Formula I forms either a three, four, six, seven oreight-membered cycloalkyl, a four to eight-membered heterocyclyl, a sixto ten-membered aryl, or a five to ten-membered heteroaryl” can beillustrated in a non-limiting matter as follows. Thus, for example, inthe case where A and M are connected such that the moiety:

shown above in Formula I forms a six-membered cycloalkyl(cyclohexyl),Formula I can be depicted as:

One with ordinary skill in the art will appreciate that similardepictions for Formula I can be arrived at when A and M shown above inthe moiety:

are connected to form a three, four, seven or eight-membered cycloalkyl,a four to eight-membered heterocyclyl, a six to ten-membered aryl, or afive to ten-membered heteroaryl.

The statement above: “alternately, (i) either R¹⁵ and R¹⁶ are connectedto each other to form a four to eight-membered cyclic structure, or R¹⁵and R¹⁹ are connected to each other to form a four to eight-memberedcyclic structure, and (ii) likewise, independently, R¹⁷ and R¹⁸ areconnected to each other to form a three to eight-membered cycloalkyl orheterocyclyl” means the following possibilities: (i) that R¹⁵ and R¹⁶are connected to form a cyclic structure while R¹⁵ and R¹⁹ are not; (ii)that R¹⁵ and R¹⁹ are connected to form a cyclic structure while R¹⁵ andR¹⁶ are not; and that (iii) R¹⁷ and R¹⁸ are independently connected toform a cyclic structure, irrespective of whether the possibilities in(i) and (ii) exist or not.

In the above-noted definitions of R, R′, R², and R³ preferred alkyl ismade of one to ten carbon atoms, preferred alkenyl or alkynyl is made oftwo to ten carbon atoms, preferred cycloalkyl is made of three to eightcarbon atoms, and preferred heteroalkyl, heteroaryl or heterocycloalkylhas one to six oxygen, nitrogen, sulfur, or phosphorus atoms.

The compounds represented by Formula I, by themselves or in combinationwith one or more other suitable agents disclosed herein, can be usefulfor treating diseases such as, for example, HCV, HIV, AIDS (AcquiredImmune Deficiency Syndrome), and related disorders, as well as formodulating the activity of hepatitis C virus (HCV) protease, preventingHCV, or ameliorating one or more symptoms of hepatitis C. Suchmodulation, treatment, prevention or amelioration can be done with theinventive compounds as well as with pharmaceutical compositions orformulations comprising such compounds. Without being limited to theory,it is believed that the HCV protease may be the NS3 or NS4a protease.The inventive compounds can inhibit such protease. They can alsomodulate the processing of hepatitis C virus (HCV) polypeptide.

DETAILED DESCRIPTION

In an embodiment, the present invention discloses compounds which arerepresented by structural Formula 1 or a pharmaceutically acceptablesalt, solvate or ester thereof, wherein the various moieties are asdefined above.

In another embodiment, R¹ is NR⁹R¹⁰, and R⁹ is H, R¹⁰ is H, or R¹⁴wherein R¹⁴ is H, alkyl, aryl, heteroalkyl, heteroaryl, cycloalkyl,alkyl-aryl, alkyl-heteroaryl, aryl-alkyl, alkenyl, alkynyl orheteroaryl-alkyl.

In another embodiment, R¹⁴ is selected from the group consisting of:

In another embodiment, R² is selected from the group consisting of thefollowing moieties:

In another embodiment, R³ is selected from the group consisting of:

wherein R³¹ is OH or O-alkyl; and

R³² is H, C(O)CH₃, C(O)OtBu or C(O)N(H)tBu.

In an additional embodiment, R³ is selected from the group consisting ofthe following moieties:

In another embodiment, Y is selected from the following moieties:

wherein R¹⁵, R¹⁶, R¹⁷, R¹⁸, and R¹⁹ can be the same or different, eachbeing independently selected from the group consisting of H, alkyl,heteroalkyl, alkenyl, heteroalkenyl, alkynyl, heteroalkynyl, cycloalkyl,heterocyclyl, aryl, and heteroaryl, or alternately, (i) either R¹⁵ andR¹⁶ are connected to form a four to eight-membered cyclic structure, orR¹⁵ and R¹⁹ are connected to form a four to eight-membered cyclicstructure, and (ii) likewise, independently, R¹⁷ and R¹⁸ are connectedto form a three to eight-membered cycloalkyl or heterocyclyl;

wherein each of said alkyl, aryl, heteroaryl, cycloalkyl or heterocyclylcan be unsubstituted or optionally independently substituted with one ormore moieties selected from the group consisting of: hydroxy, alkoxy,aryloxy, thio, alkylthio, arylthio, amino, amido, alkylamino, arylamino,alkylsulfonyl, arylsulfonyl, sulfonamido, alkylsulfonamido,arylsulfonamido, keto, carboxy, carbalkoxy, carboxamido,alkoxycarbonylamino, alkoxycarbonyloxy, alkylureido, arylureido, halo,cyano, and nitro.

In an additional embodiment, the moiety:

is selected from the group consisting of:

wherein Y³² is selected from the group consisting of:

R¹⁶ is selected from H, methyl, phenyl, benzyl; and

R¹⁵ and R¹⁹ maybe the same or different, each being independentlyselected from the following:

or alternately, the moiety:

is selected from the following moieties:

In an additional embodiment, R¹⁶ is H.

In another embodiment, the moiety:

is selected from the following structures:

In an additional embodiment, the moiety:

is selected from the following structures:

In a still additional embodiment, the moiety:

is selected from the following structures:

In a further additional embodiment, R¹ is NHR¹⁴, where R¹⁴ is selectedfrom the group consisting of:

R² is selected from the group consisting of the following moieties:

R³ is selected from the group consisting of the following moieties:

Y is selected from the group consisting of:

wherein G=NH, and the moiety,

is selected from the group consisting of:

R¹⁶═H, and

R¹⁵ and R¹⁹ can be the same or different, each being independentlyselected from the following:

or alternately, the moiety:

is represented by one of the following moieties,

and the moiety:

Yet another embodiment of the invention discloses compounds shown inTable 1, Table 1A, Table 2 and Table 3 later in this Description. Alsoshown in the Tables are the biological activities of several inventivecompounds (as Ki* values).

In an additional embodiment, this invention discloses the followingcompounds in Table 4: TABLE 4

In an additional embodiment, the present invention discloses thefollowing compounds in Table 5: TABLE 5

As used above, and throughout this disclosure, the following terms,unless otherwise indicated, shall be understood to have the followingmeanings:

“Patient” includes both human and animals.

“Mammal” means humans and other mammalian animals.

“Alkyl” means an aliphatic hydrocarbon group which may be straight orbranched and comprising about 1 to about 20 carbon atoms in the chain.Preferred alkyl groups contain about 1 to about 12 carbon atoms in thechain. More preferred alkyl groups contain about 1 to about 6 carbonatoms in the chain. Branched means that one or more lower alkyl groupssuch as methyl, ethyl or propyl, are attached to a linear alkyl chain.“Lower alkyl” means a group having about 1 to about 6 carbon atoms inthe chain which may be straight or branched. The term “substitutedalkyl” means that the alkyl group may be substituted by one or moresubstituents which may be the same or different, each substituent beingindependently selected from the group consisting of halo, alkyl, aryl,cycloalkyl, cyano, hydroxy, alkoxy, alkylthio, amino, —NH(alkyl),—NH(cycloalkyl), —N(alkyl)₂, —N(alkyl)₂, carboxy and —C(O)O-alkyl.Non-limiting examples of suitable alkyl groups include methyl, ethyl,n-propyl, isopropyl and t-butyl.

“Alkenyl” means an aliphatic hydrocarbon group containing at least onecarbon-carbon double bond and which may be straight or branched andcomprising about 2 to about 15 carbon atoms in the chain. Preferredalkenyl groups have about 2 to about 12 carbon atoms in the chain; andmore preferably about 2 to about 6 carbon atoms in the chain. Branchedmeans that one or more lower alkyl groups such as methyl, ethyl orpropyl, are attached to a linear alkenyl chain. “Lower alkenyl” meansabout 2 to about 6 carbon atoms in the chain which may be straight orbranched. The term “substituted alkenyl” means that the alkenyl groupmay be substituted by one or more substituents which may be the same ordifferent, each substituent being independently selected from the groupconsisting of halo, alkyl. aryl, cycloalkyl, cyano, alkoxy and—S(alkyl). Non-limiting examples of suitable alkenyl groups includeethenyl, propenyl, n-butenyl, 3-methylbut-2-enyl, n-pentenyl, octenyland decenyl.

“Alkynyl” means an aliphatic hydrocarbon group containing at least onecarbon-carbon triple bond and which may be straight or branched andcomprising about 2 to about 15 carbon atoms in the chain. Preferredalkynyl groups have about 2 to about 12 carbon atoms in the chain; andmore preferably about 2 to about 4 carbon atoms in the chain. Branchedmeans that one or more lower alkyl groups such as methyl, ethyl orpropyl, are attached to a linear alkynyl chain. “Lower alkynyl” meansabout 2 to about 6 carbon atoms in the chain which may be straight orbranched. Non-limiting examples of suitable alkynyl groups includeethynyl, propynyl, 2-butynyl and 3-methylbutynyl. The term “substitutedalkynyl” means that the alkynyl group may be substituted by one or moresubstituents which may be the same or different, each substituent beingindependently selected from the group consisting of alkyl, aryl andcycloalkyl.

“Aryl” means an aromatic monocyclic or multicyclic ring systemcomprising about 6 to about 14 carbon atoms, preferably about 6 to about10 carbon atoms. The aryl group can be optionally substituted with oneor more “ring system substituents” which may be the same or different,and are as defined herein. Non-limiting examples of suitable aryl groupsinclude phenyl and naphthyl.

“Heteroaryl” means an aromatic monocyclic or multicyclic ring systemcomprising about 5 to about 14 ring atoms, preferably about 5 to about10 ring atoms, in which one or more of the ring atoms is an elementother than carbon, for example nitrogen, oxygen or sulfur, alone or incombination. Preferred heteroaryls contain about 5 to about 6 ringatoms. The “heteroaryl” can be optionally substituted by one or more“ring system substituents” which may be the same or different, and areas defined herein. The prefix aza, oxa or thia before the heteroarylroot name means that at least a nitrogen, oxygen or sulfur atomrespectively, is present as a ring atom. A nitrogen atom of a heteroarylcan be optionally oxidized to the corresponding N-oxide. Non-limitingexamples of suitable heteroaryls include pyridyl, pyrazinyl, furanyl,thienyl, pyrimidinyl, pyridone (including N-substituted pyridones),isoxazolyl, isothiazolyl, oxazolyl, thiazolyl, pyrazolyl, furazanyl,pyrrolyl, pyrazolyl, triazolyl, 1,2,4-thiadiazolyl, pyrazinyl,pyridazinyl, quinoxalinyl, phthalazinyl, oxindolyl,imidazo[1,2-a]pyridinyl, imidazo[2,1-b]thiazolyl, benzofurazanyl,indolyl, azaindolyl, benzimidazolyl, benzothienyl, quinolinyl,imidazolyl, thienopyridyl, quinazolinyl, thienopyrimidyl,pyrrolopyridyl, imidazopyridyl, isoquinolinyl, benzoazaindolyl,1,2,4-triazinyl, benzothiazolyl and the like. The term “heteroaryl” alsorefers to partially saturated heteroaryl moieties such as, for example,tetrahydroisoquinolyl, tetrahydroquinolyl and the like.

“Aralkyl” or “arylalkyl” means an aryl-alkyl-group in which the aryl andalkyl are as previously described. Preferred aralkyls comprise a loweralkyl group. Non-limiting examples of suitable aralkyl groups includebenzyl, 2-phenethyl and naphthalenylmethyl. The bond to the parentmoiety is through the alkyl.

“Alkylaryl” means an alkyl-aryl-group in which the alkyl and aryl are aspreviously described. Preferred alkylaryls comprise a lower alkyl group.Non-limiting example of a suitable alkylaryl group is tolyl. The bond tothe parent moiety is through the aryl.

“Cycloalkyl” means a non-aromatic mono- or multicyclic ring systemcomprising about 3 to about 10 carbon atoms, preferably about 5 to about10 carbon atoms. Preferred cycloalkyl rings contain about 5 to about 7ring atoms. The cycloalkyl can be optionally substituted with one ormore “ring system substituents” which may be the same or different, andare as defined above. Non-limiting examples of suitable monocycliccycloalkyls include cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyland the like. Non-limiting examples of suitable multicyclic cycloalkylsinclude 1-decalinyl, norbornyl, adamantyl and the like, as well aspartially saturated species such as, for example, indanyl,tetrahydronaphthyl and the like.

“Halogen” or “halo” means fluorine, chlorine, bromine, or iodine.Preferred are fluorine, chlorine and bromine.

“Ring system substituent” means a substituent attached to an aromatic ornon-aromatic ring system which, for example, replaces an availablehydrogen on the ring system. Ring system substituents may be the same ordifferent, each being independently selected from the group consistingof alkyl, alkenyl, alkynyl, aryl, heteroaryl, aralkyl, alkylaryl,heteroaralkyl, heteroarylalkenyl, heteroarylalkynyl, alkylheteroaryl,hydroxy, hydroxyalkyl, alkoxy, aryloxy, aralkoxy, acyl, aroyl, halo,nitro, cyano, carboxy, alkoxycarbonyl, aryloxycarbonyl,aralkoxycarbonyl, alkylsulfonyl, arylsulfonyl, heteroarylsulfonyl,alkylthio, arylthio, heteroarylthio, aralkylthio, heteroaralkylthio,cycloalkyl, heterocyclyl, —C(═N—CN)—NH₂, —C(═NH)—NH₂, —C(═NH)—NH(alkyl),Y₁Y₂N—, Y₁Y₂N-alkyl-, Y₁Y₂NC(O)—, Y₁Y₂NSO₂— and —SO₂NY₁Y₂, wherein Y₁and Y₂ can be the same or different and are independently selected fromthe group consisting of hydrogen, alkyl, aryl, cycloalkyl, and aralkyl.“Ring system substituent” may also mean a single moiety whichsimultaneously replaces two available hydrogens on two adjacent carbonatoms (one H on each carbon) on a ring system. Examples of such moietyare methylene dioxy, ethylenedioxy, —C(CH₃)₂— and the like which formmoieties such as, for example:

“Heterocyclyl” means a non-aromatic saturated monocyclic or multicyclicring system comprising about 3 to about 10 ring atoms, preferably about5 to about 10 ring atoms, in which one or more of the atoms in the ringsystem is an element other than carbon, for example nitrogen, oxygen orsulfur, alone or in combination. There are no adjacent oxygen and/orsulfur atoms present in the ring system. Preferred heterocyclyls containabout 5 to about 6 ring atoms. The prefix aza, oxa or thia before theheterocyclyl root name means that at least a nitrogen, oxygen or sulfuratom respectively is present as a ring atom. Any —NH in a heterocyclylring may exist protected such as, for example, as an —N(Boc), —N(CBz),—N(Tos) group and the like; such protections are also considered part ofthis invention. The heterocyclyl can be optionally substituted by one ormore “ring system substituents” which may be the same or different, andare as defined herein. The nitrogen or sulfur atom of the heterocyclylcan be optionally oxidized to the corresponding N-oxide, S-oxide orS,S-dioxide. Non-limiting examples of suitable monocyclic heterocyclylrings include piperidyl, pyrrolidinyl, piperazinyl, morpholinyl,thiomorpholinyl, thiazolidinyl, 1,4-dioxanyl, tetrahydrofuranyl,tetrahydrothiophenyl, lactam, lactone, and the like.

It should be noted that in hetero-atom containing ring systems of thisinvention, there are no hydroxyl groups on carbon atoms adjacent to a N,O or S, as well as there are no N or S groups on carbon adjacent toanother heteroatom. Thus, for example, in the ring:

there is no —OH attached directly to carbons marked 2 and 5.

It should also be noted that tautomeric forms such as, for example, themoieties:

are considered equivalent in certain embodiments of this invention.

“Alkynylalkyl” means an alkynyl-alkyl-group in which the alkynyl andalkyl are as previously described. Preferred alkynylalkyls contain alower alkynyl and a lower alkyl group. The bond to the parent moiety isthrough the alkyl. Non-limiting examples of suitable alkynylalkyl groupsinclude propargylmethyl.

“Heteroaralkyl” means a heteroaryl-alkyl-group in which the heteroaryland alkyl are as previously described. Preferred heteroaralkyls containa lower alkyl group. Non-limiting examples of suitable aralkyl groupsinclude pyridylmethyl, and quinolin-3-ylmethyl. The bond to the parentmoiety is through the alkyl.

“Hydroxyalkyl” means a HO-alkyl-group in which alkyl is as previouslydefined. Preferred hydroxyalkyls contain lower alkyl. Non-limitingexamples of suitable hydroxyalkyl groups include hydroxymethyl and2-hydroxyethyl.

“Acyl” means an H—C(O)—, alkyl-C(O)— or cycloalkyl-C(O)—, group in whichthe various groups are as previously described. The bond to the parentmoiety is through the carbonyl. Preferred acyls contain a lower alkyl.Non-limiting examples of suitable acyl groups include formyl, acetyl andpropanoyl.

“Aroyl” means an aryl-C(O)-group in which the aryl group is aspreviously described. The bond to the parent moiety is through thecarbonyl. Non-limiting examples of suitable groups include benzoyl and1-naphthoyl.

“Alkoxy” means an alkyl-O-group in which the alkyl group is aspreviously described. Non-limiting examples of suitable alkoxy groupsinclude methoxy, ethoxy, n-propoxy, isopropoxy and n-butoxy. The bond tothe parent moiety is through the ether oxygen.

“Aryloxy” means an aryl-O-group in which the aryl group is as previouslydescribed. Non-limiting examples of suitable aryloxy groups includephenoxy and naphthoxy. The bond to the parent moiety is through theether oxygen.

“Aralkyloxy” means an aralkyl-O-group in which the aralkyl group is aspreviously described. Non-limiting examples of suitable aralkyloxygroups include benzyloxy and 1- or 2-naphthalenemethoxy. The bond to theparent moiety is through the ether oxygen.

“Alkylthio” means an alkyl-S-group in which the alkyl group is aspreviously described. Non-limiting examples of suitable alkylthio groupsinclude methylthio and ethylthio. The bond to the parent moiety isthrough the sulfur.

“Arylthio” means an aryl-S-group in which the aryl group is aspreviously described. Non-limiting examples of suitable arylthio groupsinclude phenylthio and naphthylthio. The bond to the parent moiety isthrough the sulfur.

“Aralkylthio” means an aralkyl-S-group in which the aralkyl group is aspreviously described. Non-limiting example of a suitable aralkylthiogroup is benzylthio. The bond to the parent moiety is through thesulfur.

“Alkoxycarbonyl” means an alkyl-O—CO-group. Non-limiting examples ofsuitable alkoxycarbonyl groups include methoxycarbonyl andethoxycarbonyl. The bond to the parent moiety is through the carbonyl.

“Aryloxycarbonyl” means an aryl-O—C(O)-group. Non-limiting examples ofsuitable aryloxycarbonyl groups include phenoxycarbonyl andnaphthoxycarbonyl. The bond to the parent moiety is through thecarbonyl.

“Aralkoxycarbonyl” means an aralkyl-O—C(O)-group. Non-limiting exampleof a suitable aralkoxycarbonyl group is benzyloxycarbonyl. The bond tothe parent moiety is through the carbonyl.

“Alkylsulfonyl” means an alkyl-S(O₂)-group. Preferred groups are thosein which the alkyl group is lower alkyl. The bond to the parent moietyis through the sulfonyl.

“Arylsulfonyl” means an aryl-S(O₂)-group. The bond to the parent moietyis through the sulfonyl.

The term “substituted” means that one or more hydrogens on thedesignated atom is replaced with a selection from the indicated group,provided that the designated atom's normal valency under the existingcircumstances is not exceeded, and that the substitution results in astable compound. Combinations of substituents and/or variables arepermissible only if such combinations result in stable compounds. By“stable compound’ or “stable structure” is meant a compound that issufficiently robust to survive isolation to a useful degree of purityfrom a reaction mixture, and formulation into an efficacious therapeuticagent.

The term “one or more” or “at least one”, when indicating the number ofsubstituents, compounds, combination agents and the like, refers to atleast one, and up to the maximum number of chemically and physicallypermissible, substituents, compounds, combination agents and the like,that are present or added, depending on the context. Such techniques andknowledge are well known within the skills of the concerned artisan.

The term “optionally substituted” means optional substitution with thespecified groups, radicals or moieties.

The term “isolated” or “in isolated form” for a compound refers to thephysical state of said compound after being isolated from a syntheticprocess or natural source or combination thereof. The term “purified” or“in purified form” for a compound refers to the physical state of saidcompound after being obtained from a purification process or processesdescribed herein or well known to the skilled artisan, in sufficientpurity to be characterizable by standard analytical techniques describedherein or well known to the skilled artisan.

It should also be noted that any heteroatom with unsatisfied valences inthe text, schemes, examples and Tables herein is assumed to have thehydrogen atom(s) to satisfy the valences.

When a functional group in a compound is termed “protected”, this meansthat the group is in modified form to preclude undesired side reactionsat the protected site when the compound is subjected to a reaction.Suitable protecting groups will be recognized by those with ordinaryskill in the art as well as by reference to standard textbooks such as,for example, T. W. Greene et al, Protective Groups in organic Synthesis(1991), Wiley, N.Y.

When any variable (e.g., aryl, heterocycle, R², etc.) occurs more thanone time in any constituent or in Formula 1, its definition on eachoccurrence is independent of its definition at every other occurrence.

As used herein, the term “composition” is intended to encompass aproduct comprising the specified ingredients in the specified amounts,as well as any product which results, directly or indirectly, fromcombination of the specified ingredients in the specified amounts.

Prodrugs and solvates of the compounds of the invention are alsocontemplated herein. The term “prodrug”, as employed herein, denotes acompound that is a drug precursor which, upon administration to asubject, undergoes chemical conversion by metabolic or chemicalprocesses to yield a compound of Formula 1 or a salt and/or solvatethereof. A discussion of prodrugs is provided in T. Higuchi and V.Stella, Pro-drugs as Novel Delivery Systems (1987) 14 of the A.C.S.Symposium Series, and in Bioreversible Carriers in Drug Design, (1987)Edward B. Roche, ed., American Pharmaceutical Association and PergamonPress, both of which are incorporated herein by reference thereto.

“Solvate” means a physical association of a compound of this inventionwith one or more solvent molecules. This physical association involvesvarying degrees of ionic and covalent bonding, including hydrogenbonding. In certain instances the solvate will be capable of isolation,for example when one or more solvent molecules are incorporated in thecrystal lattice of the crystalline solid. “Solvate” encompasses bothsolution-phase and isolatable solvates. Non-limiting examples ofsuitable solvates include ethanolates, methanolates, and the like.“Hydrate” is a solvate wherein the solvent molecule is H₂O.

“Effective amount” or “therapeutically effective amount” is meant todescribe an amount of compound or a composition of the present inventioneffective in inhibiting the CDK(s) and thus producing the desiredtherapeutic, ameliorative, inhibitory or preventative effect.

The compounds of Formula 1 can form salts which are also within thescope of this invention. Reference to a compound of Formula 1 herein isunderstood to include reference to salts thereof, unless otherwiseindicated. The term “salt(s)”, as employed herein, denotes acidic saltsformed with inorganic and/or organic acids, as well as basic saltsformed with inorganic and/or organic bases. In addition, when a compoundof Formula 1 contains both a basic moiety, such as, but not limited to apyridine or imidazole, and an acidic moiety, such as, but not limited toa carboxylic acid, zwitterions (“inner salts”) may be formed and areincluded within the term “salt(s)” as used herein. Pharmaceuticallyacceptable (i.e., non-toxic, physiologically acceptable) salts arepreferred, although other salts are also useful. Salts of the compoundsof the Formula 1 may be formed, for example, by reacting a compound ofFormula 1 with an amount of acid or base, such as an equivalent amount,in a medium such as one in which the salt precipitates or in an aqueousmedium followed by lyophilization.

Exemplary acid addition salts include acetates, ascorbates, benzoates,benzenesulfonates, bisulfates, borates, butyrates, citrates,camphorates, camphorsulfonates, fumarates, hydrochlorides,hydrobromides, hydroiodides, lactates, maleates, methanesulfonates,naphthalenesulfonates, nitrates, oxalates, phosphates, propionates,salicylates, succinates, sulfates, tartarates, thiocyanates,toluenesulfonates (also known as tosylates,) and the like. Additionally,acids which are generally considered suitable for the formation ofpharmaceutically useful salts from basic pharmaceutical compounds arediscussed, for example, by P. Stahl et al, Camille G. (eds.) Handbook ofPharmaceutical Salts. Properties, Selection and Use. (2002) Zurich:Wiley-VCH; S. Berge et al, Journal of Pharmaceutical Sciences (1977)66(1) 1-19; P. Gould, International J. of Pharmaceutics (1986) 33201-217; Anderson et al, The Practice of Medicinal Chemistry (1996),Academic Press, New York; and in The Orange Book (Food & DrugAdministration, Washington, D.C. on their website). These disclosuresare incorporated herein by reference thereto.

Exemplary basic salts include ammonium salts, alkali metal salts such assodium, lithium, and potassium salts, alkaline earth metal salts such ascalcium and magnesium salts, salts with organic bases (for example,organic amines) such as dicyclohexylamines, t-butyl amines, and saltswith amino acids such as arginine, lysine and the like. Basicnitrogen-containing groups may be quarternized with agents such as loweralkyl halides (e.g. methyl, ethyl, and butyl chlorides, bromides andiodides), dialkyl sulfates (e.g. dimethyl, diethyl, and dibutylsulfates), long chain halides (e.g. decyl, lauryl, and stearylchlorides, bromides and iodides), aralkyl halides (e.g. benzyl andphenethyl bromides), and others.

All such acid salts and base salts are intended to be pharmaceuticallyacceptable salts within the scope of the invention and all acid and basesalts are considered equivalent to the free forms of the correspondingcompounds for purposes of the invention.

Pharmaceutically acceptable esters of the present compounds include thefollowing groups: (1) carboxylic acid esters obtained by esterificationof the hydroxy groups, in which the non-carbonyl moiety of thecarboxylic acid portion of the ester grouping is selected from straightor branched chain alkyl (for example, acetyl, n-propyl, t-butyl, orn-butyl), alkoxyalkyl (for example, methoxymethyl), aralkyl (forexample, benzyl), aryloxyalkyl (for example, phenoxymethyl), aryl (forexample, phenyl optionally substituted with, for example, halogen,C₁₋₄alkyl, or C₁₋₄alkoxy or amino); (2) sulfonate esters, such as alkyl-or aralkylsulfonyl (for example, methanesulfonyl); (3) amino acid esters(for example, L-valyl or L-isoleucyl); (4) phosphonate esters and (5)mono-, di- or triphosphate esters. The phosphate esters may be furtheresterified by, for example, a C₁₋₂₀ alcohol or reactive derivativethereof, or by a 2,3-di(C₆₋₂₄)acyl glycerol.

Compounds of Formula 1, and salts, solvates, esters and prodrugsthereof, may exist in their tautomeric form (for example, as an amide orimino ether). All such tautomeric forms are contemplated herein as partof the present invention.

All stereoisomers (for example, geometric isomers, optical isomers, andthe like) of the present compounds (including those of the salts,solvates and prodrugs of the compounds as well as the salts and solvatesof the prodrugs), such as those which may exist due to asymmetriccarbons on various substituents, including enantiomeric forms (which mayexist even in the absence of asymmetric carbons), rotameric forms,atropisomers, and diastereomeric forms, are contemplated within thescope of this invention, as are positional isomers (such as, forexample, 4-pyridyl and 3-pyridyl). Individual stereoisomers of thecompounds of the invention may, for example, be substantially free ofother isomers, or may be admixed, for example, as racemates or with allother, or other selected, stereoisomers. The chiral centers of thepresent invention can have the S or R configuration as defined by theIUPAC 1974 Recommendations. The use of the terms “salt”, “solvate”“prodrug” and the like, is intended to equally apply to the salt,solvate and prodrug of enantiomers, stereoisomers, rotamers, tautomers,positional isomers, racemates or prodrugs of the inventive compounds.

Polymorphic forms of the compounds of Formula I, and of the salts,solvates, esters and prodrugs of the compounds of Formula I, areintended to be included in the present invention.

It is to be understood that the utility of the compounds of Formula 1for the therapeutic applications discussed herein is applicable to eachcompound by itself or to the combination or combinations of one or morecompounds of Formula 1 as illustrated, for example, in the nextimmediate paragraph. The same understanding also applies topharmaceutical composition(s) comprising such compound or compounds andmethod(s) of treatment involving such compound or compounds.

The compounds according to the invention can have pharmacologicalproperties; in particular, the compounds of Formula 1 can be inhibitorsof HCV protease, each compound by itself or one or more compounds ofFormula 1 can be combined with one or more compounds selected fromwithin Formula 1. The compound(s) can be useful for treating diseasessuch as, for example, HCV, HIV, (AIDS, Acquired Immune DeficiencySyndrome), and related disorders, as well as for modulating the activityof hepatitis C virus (HCV) protease, preventing HCV, or ameliorating oneor more symptoms of hepatitis C.

The compounds of Formula 1 may be used for the manufacture of amedicament to treat disorders associated with the HCV protease, forexample, the method comprising bringing into intimate contact a compoundof Formula 1 and a pharmaceutically acceptable carrier.

In another embodiment, this invention provides pharmaceuticalcompositions comprising the inventive compound or compounds as an activeingredient. The pharmaceutical compositions generally additionallycomprise at least one pharmaceutically acceptable carrier diluent,excipient or carrier (collectively referred to herein as carriermaterials). Because of their HCV inhibitory activity, suchpharmaceutical compositions possess utility in treating hepatitis C andrelated disorders.

In yet another embodiment, the present invention discloses methods forpreparing pharmaceutical compositions comprising the inventive compoundsas an active ingredient. In the pharmaceutical compositions and methodsof the present invention, the active ingredients will typically beadministered in admixture with suitable carrier materials suitablyselected with respect to the intended form of administration, i.e. oraltablets, capsules (either solid-filled, semi-solid filled or liquidfilled), powders for constitution, oral gels, elixirs, dispersiblegranules, syrups, suspensions, and the like, and consistent withconventional pharmaceutical practices. For example, for oraladministration in the form of tablets or capsules, the active drugcomponent may be combined with any oral non-toxic pharmaceuticallyacceptable inert carrier, such as lactose, starch, sucrose, cellulose,magnesium stearate, dicalcium phosphate, calcium sulfate, talc,mannitol, ethyl alcohol (liquid forms) and the like. Moreover, whendesired or needed, suitable binders, lubricants, disintegrating agentsand coloring agents may also be incorporated in the mixture. Powders andtablets may be comprised of from about 5 to about 95 percent inventivecomposition.

Suitable binders include starch, gelatin, natural sugars, cornsweeteners, natural and synthetic gums such as acacia, sodium alginate,carboxymethylcellulose, polyethylene glycol and waxes. Among thelubricants there may be mentioned for use in these dosage forms, boricacid, sodium benzoate, sodium acetate, sodium chloride, and the like.Disintegrants include starch, methylcellulose, guar gum and the like.

Sweetening and flavoring agents and preservatives may also be includedwhere appropriate. Some of the terms noted above, namely disintegrants,diluents, lubricants, binders and the like, are discussed in more detailbelow.

Additionally, the compositions of the present invention may beformulated in sustained release form to provide the rate controlledrelease of any one or more of the components or active ingredients tooptimize the therapeutic effects, i.e. HCV inhibitory activity and thelike. Suitable dosage forms for sustained release include layeredtablets containing layers of varying disintegration rates or controlledrelease polymeric matrices impregnated with the active components andshaped in tablet form or capsules containing such impregnated orencapsulated porous polymeric matrices.

Liquid form preparations include solutions, suspensions and emulsions.As an example may be mentioned water or water-propylene glycol solutionsfor parenteral injections or addition of sweeteners and pacifiers fororal solutions, suspensions and emulsions. Liquid form preparations mayalso include solutions for intranasal administration.

Aerosol preparations suitable for inhalation may include solutions andsolids in powder form, which may be in combination with apharmaceutically acceptable carrier such as inert compressed gas, e.g.nitrogen.

For preparing suppositories, a low melting wax such as a mixture offatty acid glycerides such as cocoa buffer is first melted, and theactive ingredient is dispersed homogeneously therein by stirring orsimilar mixing. The molten homogeneous mixture is then poured intoconvenient sized molds, allowed to cool and thereby solidify.

Also included are solid form preparations which are intended to beconverted, shortly before use, to liquid form preparations for eitheroral or parenteral administration. Such liquid forms include solutions,suspensions and emulsions.

The compounds of the invention may also be deliverable transdermally.The transdermal compositions may take the form of creams, lotions,aerosols and/or emulsions and can be included in a transdermal patch ofthe matrix or reservoir type as are conventional in the art for thispurpose.

The compounds of the invention may also be administered orally,intravenously, intranasally or subcutaneously.

The compounds of the invention may also comprise preparations which arein a unit dosage form. In such form, the preparation is subdivided intosuitably sized unit doses containing appropriate quantities of theactive components, e.g., an effective amount to achieve the desiredpurpose.

The quantity of the inventive active composition in a unit dose ofpreparation may be generally varied or adjusted from about 1.0 milligramto about 1,000 milligrams, preferably from about 1.0 to about 950milligrams, more preferably from about 1.0 to about 500 milligrams, andtypically from about 1 to about 250 milligrams, according to theparticular application. The actual dosage employed may be varieddepending upon the patient's age, sex, weight and severity of thecondition being treated. Such techniques are well known to those skilledin the art.

Generally, the human oral dosage form containing the active ingredientscan be administered 1 or 2 times per day. The amount and frequency ofthe administration will be regulated according to the judgment of theattending clinician. A generally recommended daily dosage regimen fororal administration may range from about 1.0 milligram to about 1,000milligrams per day, in single or divided doses.

Some useful terms are described below:

Capsule—refers to a special container or enclosure made of methylcellulose, polyvinyl alcohols, or denatured gelatins or starch forholding or containing compositions comprising the active ingredients.Hard shell capsules are typically made of blends of relatively high gelstrength bone and pork skin gelatins. The capsule itself may containsmall amounts of dyes, opaquing agents, plasticizers and preservatives.

Tablet—refers to a compressed or molded solid dosage form containing theactive ingredients with suitable diluents. The tablet can be prepared bycompression of mixtures or granulations obtained by wet granulation, drygranulation or by compaction.

Oral gel—refers to the active ingredients dispersed or solubilized in ahydrophillic semi-solid matrix.

Powder for constitution refers to powder blends containing the activeingredients and suitable diluents which can be suspended in water orjuices.

Diluent—refers to substances that usually make up the major portion ofthe composition or dosage form. Suitable diluents include sugars such aslactose, sucrose, mannitol and sorbitol; starches derived from wheat,corn, rice and potato; and celluloses such as microcrystallinecellulose. The amount of diluent in the composition can range from about10 to about 90% by weight of the total composition, preferably fromabout 25 to about 75%, more preferably from about 30 to about 60% byweight, even more preferably from about 12 to about 60%.

Disintegrant—refers to materials added to the composition to help itbreak apart (disintegrate) and release the medicaments. Suitabledisintegrants include starches; “cold water soluble” modified starchessuch as sodium carboxymethyl starch; natural and synthetic gums such aslocust bean, karaya, guar, tragacanth and agar; cellulose derivativessuch as methylcellulose and sodium carboxymethylcellulose;microcrystalline celluloses and cross-linked microcrystalline cellulosessuch as sodium croscarmellose; alginates such as alginic acid and sodiumalginate; clays such as bentonites; and effervescent mixtures. Theamount of disintegrant in the composition can range from about 2 toabout 15% by weight of the composition, more preferably from about 4 toabout 10% by weight.

Binder—refers to substances that bind or “glue” powders together andmake them cohesive by forming granules, thus serving as the “adhesive”in the formulation. Binders add cohesive strength already available inthe diluent or bulking agent. Suitable binders include sugars such assucrose; starches derived from wheat, corn rice and potato; natural gumssuch as acacia, gelatin and tragacanth; derivatives of seaweed such asalginic acid, sodium alginate and ammonium calcium alginate; cellulosicmaterials such as methylcellulose and sodium carboxymethylcellulose andhydroxypropylmethylcellulose; polyvinylpyrrolidone; and inorganics suchas magnesium aluminum silicate. The amount of binder in the compositioncan range from about 2 to about 20% by weight of the composition, morepreferably from about 3 to about 10% by weight, even more preferablyfrom about 3 to about 6% by weight.

Lubricant—refers to a substance added to the dosage form to enable thetablet, granules, etc. after it has been compressed, to release from themold or die by reducing friction or wear. Suitable lubricants includemetallic stearates such as magnesium stearate, calcium stearate orpotassium stearate; stearic acid; high melting point waxes; and watersoluble lubricants such as sodium chloride, sodium benzoate, sodiumacetate, sodium oleate, polyethylene glycols and d′l-leucine. Lubricantsare usually added at the very last step before compression, since theymust be present on the surfaces of the granules and in between them andthe parts of the tablet press. The amount of lubricant in thecomposition can range from about 0.2 to about 5% by weight of thecomposition, preferably from about 0.5 to about 2%, more preferably fromabout 0.3 to about 1.5% by weight.

Glident—material that prevents caking and improve the flowcharacteristics of granulations, so that flow is smooth and uniform.Suitable glidents include silicon dioxide and talc. The amount ofglident in the composition can range from about 0.1% to about 5% byweight of the total composition, preferably from about 0.5 to about 2%by weight.

Coloring agents—excipients that provide coloration to the composition orthe dosage form. Such excipients can include food grade dyes and foodgrade dyes adsorbed onto a suitable adsorbent such as clay or aluminumoxide. The amount of the coloring agent can vary from about 0.1 to about5% by weight of the composition, preferably from about 0.1 to about 1%.

Bioavailability—refers to the rate and extent to which the active drugingredient or therapeutic moiety is absorbed into the systemiccirculation from an administered dosage form as compared to a standardor control.

Conventional methods for preparing tablets are known. Such methodsinclude dry methods such as direct compression and compression ofgranulation produced by compaction, or wet methods or other specialprocedures. Conventional methods for making other forms foradministration such as, for example, capsules, suppositories and thelike are also well known.

Another embodiment of the invention discloses the use of the inventivecompounds or pharmaceutical compositions disclosed above for treatmentof diseases such as, for example, hepatitis C and the like. The methodcomprises administering a therapeutically effective amount of theinventive compound or pharmaceutical composition to a patient havingsuch a disease or diseases and in need of such a treatment.

In yet another embodiment, the compounds of the invention may be usedfor the treatment of HCV in humans in monotherapy mode or in acombination therapy (e.g., dual combination, triple combination etc.)mode such as, for example, in combination with antiviral and/orimmunomodulatory agents. Examples of such antiviral and/orimmunomodulatory agents include Ribavirin (from Schering-PloughCorporation, Madison, N.J.) and Levovirin™ (from ICN Pharmaceuticals,Costa Mesa, Calif.), VP 50406™ (from Viropharma, Incorporated, Exton,Pa.), ISIS 14803™ (from ISIS Pharmaceuticals, Carlsbad, Calif.),Heptazyme™ (from Ribozyme Pharmaceuticals, Boulder, Colo.), VX 497™(from Vertex Pharmaceuticals, Cambridge, Mass.), Thymosin™ (fromSciClone Pharmaceuticals, San Mateo, Calif.), Maxamine (MaximPharmaceuticals, San Diego, Calif.), mycophenolate mofetil (fromHoffman-LaRoche, Nutley, N.J.), interferon (such as, for example,interferon-alpha, PEG-interferon alpha conjugates) and the like.“PEG-interferon alpha conjugates” are interferon alpha moleculescovalently attached to a PEG molecule. Illustrative PEG-interferon alphaconjugates include interferon alpha-2a (Roferon™, from Hoffman La-Roche,Nutley, N.J.) in the form of pegylated interferon alpha-2a (e.g., assold under the trade name Pegasys™), interferon alpha-2b (Intron™, fromSchering-Plough Corporation) in the form of pegylated interferonalpha-2b (e.g., as sold under the trade name PEG-Intron™), interferonalpha-2c (Berofor Alpha™, from Boehringer Ingelheim, Ingelheim, Germany)or consensus interferon as defined by determination of a consensussequence of naturally occurring interferon alphas (Infergen™, fromAmgen, Thousand Oaks, Calif.).

As stated earlier, the invention includes tautomers, rotamers,enantiomers and other stereoisomers of the inventive compounds also.Thus, as one skilled in the art appreciates, some of the inventivecompounds may exist in suitable isomeric forms. Such variations arecontemplated to be within the scope of the invention.

Another embodiment of the invention discloses a method of making thecompounds disclosed herein. The compounds may be prepared by severaltechniques known in the art. Illustrative procedures are outlined in thefollowing reaction schemes. The illustrations should not be construed tolimit the scope of the invention which is defined in the appendedclaims. Alternative mechanistic pathways and analogous structures willbe apparent to those skilled in the art.

It is to be understood that while the following illustrative schemesdescribe the preparation of a few representative inventive compounds,suitable substitution of any of both the natural and unnatural aminoacids will result in the formation of the desired compounds based onsuch substitution. Such variations are contemplated to be within thescope of the invention.

For the procedures described below, the following abbreviations areused:

ABBREVIATIONS

Abbreviations which are used in the descriptions of the schemes,preparations and the examples that follow are:

-   THF: Tetrahydrofuran-   DMF: N,N-Dimethylformamide-   EtOAc: Ethyl acetate-   AcOH: Acetic acid-   HOOBt: 3-Hydroxy-1,2,3-benzotriazin-4(3H)-one-   EDCI: 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride-   NMM: N-Methylmorpholine-   ADDP: 1,1′-(Azodicarbobyl)dipiperidine-   DEAD: Diethylazodicarboxylate-   MeOH: Methanol-   EtOH: Ethanol-   Et₂O: Diethyl ether-   DMSO: Dimethylsulfoxide-   HOBt: N-Hydroxybenzotriazole-   PyBrOP: Bromo-tris-pyrrolidinophosphonium hexafluorophosphate-   DCM: Dichloromethane-   DCC: 1,3-Dicyclohexylcarbodiimide-   TEMPO: 2,2,6,6-Tetramethyl-1-piperidinyloxy-   Phg: Phenylglycine-   Chg: Cyclohexylglycine-   Bn: Benzyl-   Bzl: Benzyl-   Et: Ethyl-   Ph: Phenyl-   iBoc: isobutoxycarbonyl-   iPr: isopropyl-   ^(t)Bu or Bu^(t): tert-Butyl-   Boc: tert-Butyloxycarbonyl-   Cbz: Benzyloxycarbonyl-   Cp: Cylcopentyldienyl-   Ts: p-toluenesulfonyl-   Me: Methyl-   HATU: O-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium    hexafluorophosphate-   DMAP: 4-N,N-Dimethylaminopyridine-   BOP: Benzotriazol-1-yl-oxy-tris(dimethylamino)hexafluorophosphate-   PCC: Pyridiniumchlorochromate

General Schemes for Preparation of Target Compounds

Compounds of the present invention were synthesized using the generalschemes (Methods A-E) described below.

Method A:

Deprotection of the N-Boc functionality of 1.01 under acidic conditionsprovided the hydrochloride salt 1.02 which was subsequently coupled withN-Boc-tert-leucine under peptide coupling methodology to afford 1.03.N-Boc deprotection followed by treatment with appropriate isocyanategave the urea 1.05. Hydrolysis of the methyl ester provided the acid1.06. Peptide coupling of the acid 1.06 with the appropriate P₁—P′primary amide moiety afforded the hydroxyl amide 1.07. Oxidation(Moffatt or related process—T. T. Tidwell, Synthesis, 1990, 857; orDess-Martin's—J. Org. Chem., 1983, 48, 4155) resulted in the targetcompound 1.08.

Method B

Peptide coupling of the acid 1.06 with the appropriate P₁—P′ secondaryamide moiety afforded the hydroxyl amide 1.09. Oxidation (Moffatt orDess-Martin's) resulted in the target compound 1.10.

Method C

In another variation, peptide coupling of the N-Boc-P₂—P₃-acid 1.17 withthe appropriate P₁—P′ amide moiety afforded the hydroxyl amide 1.11.Oxidation (Moffatt or Dess-Martin's) resulted in the keto amide 1.12.Deprotection of the N-Boc functionality gave the hydrochloride salt1.13. Treatment with a suitable isocyanate (or isocyanate equivalent)resulted in the target compound 1.14.

Method D

In yet another variation, the hydrochloride salt 1.13 was converted tothe 4-nitrophenyl carbamate 1.15 by reaction with 4-nitrophenylchloroformate. Subsequent treatment with an amine (or aminehydrochloride salt) of choice provided the target compound 1.14.

Method E

In yet another variation, the dipeptide hydrochloride salt 1.03 wasconverted to the 4-nitrophenyl carbamate as described above. Treatmentwith an amine (or amine hydrochloride salt) of choice provided the ureaderivative 1.05. Hydrolysis and further elaboration as described inMethods A/B provided the target compounds 1.14.

Preparation of Intermediates:Preparation of P₁—P′ Moieties:Preparation of Intermediates 10.11 and 10.12:Step 1:

A stirred solution of ketimine 10.01 (50 g, 187.1 mmol) under N₂ in dryTHF (400 mL) was cooled to −78° C. and treated with 1 M solution ofK—^(t)BuO (220 mL, 1.15 equiv.) in THF. The reaction mixture was warmedto 0° C. and stirred for 1 h and treated with bromomethyl cyclobutane(28 mL, 249 mmol). The reaction mixture was stirred at room temperaturefor 48 h and concentrated in vacuo. The residue was dissolved in Et₂O(300 mL) and treated with aq. HCl (2 M, 300 mL) The resulting solutionwas stirred at room temperature for 5 h and extracted with Et₂O (1 L).The aqueous layer was made basic to pH ˜12-14 with NaOH (50% aq.) andextracted with CH₂Cl₂ (3×300 mL). The combined organic layers were dried(MgSO₄), filtered, and concentrated to give the pure amine (10.02, 18 g)as a colorless oil.Step 2

A solution of the amine 10.02 (18 g, 105.2 mmol) at 0° C. in CH₂Cl₂ (350mL) was treated with di-tert-butyldicarbonate (23 g, 105.4 mmol) andstirred at rt. for 12 h. After the completion of the reaction (TLC), thereaction mixture was concentrated in vacuo and the residue was dissolvedin THF/H₂O (200 ml, 1:1) and treated with LiOH.H₂O (6.5 g, 158.5 mmol)and stirred at room temperature for 3 h. The reaction mixture wasconcentrated and the basic aqueous layer was extracted with Et₂O. Theaqueous layer was acidified with conc. HCl to pH˜1-2 and extracted withCH₂Cl₂. The combined organic layers were dried (MgSO₄), filtered, andconcentrated in vacuo to yield 10.03 as a colorless viscous oil whichwas used for the next step without any further purification.

Step 3

A solution of the acid 10.03 (15.0 g, 62 mmol) in CH₂Cl₂ (250 mL) wastreated with BOP reagent (41.1 g, 93 mmol), N-methyl morpholine (27 mL),N,O-dimethyl hydroxylamine hydrochloride (9.07 g, 93 mmol) and stirredovernight at rt. The reaction mixture was diluted with 1 N aq. HCl (250mL), and the layers were separated and the aqueous layer was extractedwith CH₂Cl₂ (3×300 ml). The combined organic layers were dried (MgSO₄),filtered and concentrated in vacuo and purified by chromatography (SiO₂,EtOAc/Hex 2:3) to yield the amide 10.04 (15.0 g) as a colorless solid.Step 4

A solution of the amide 10.04 (15 g, 52.1 mmol) in dry THF (200 mL) wastreated dropwise with a solution of LiAlH₄ (1M, 93 mL, 93 mmol) at 0° C.The reaction mixture was stirred at room temperature for 1 h andcarefully quenched at 0° C. with a solution of KHSO₄ (10% aq.) andstirred for 0.5 h. The reaction mixture was diluted with aq. HCl (1 M,150 mL) and extracted with CH₂Cl₂ (3×200 mL), The combined organiclayers were washed with aq. HCl (1 M), saturated NaHCO₃, brine, anddried (MgSO₄). The mixture was filtered and concentrated in vacuo toyield 10.05 as a viscous colorless oil (14 g).Step 5

A solution of the aldehyde 10.05 (14 g, 61.6 mmol) in CH₂Cl₂ (50 mL),was treated with Et₃N (10.73 mL, 74.4 mmol), and acetone cyanohydrin(10.86 g, 127.57 mmol) and stirred at room temperature for 24 hrs. Thereaction mixture was concentrated in vacuo and diluted with aq. HCl (1M, 200 mL) and extracted into CH₂Cl₂ (3×200 mL). The combined organiclayer were washed with H₂O, brine, dried (MgSO₄), filtered, concentratedin vacuo and purified by chromatography (SiO₂, EtOAc/Hex 1:4) to yield10.06 (10.3 g) as a colorless liquidStep 6

Methanol saturated with HCl*, prepared by bubbling HCl gas through CH₃OH(700 ml) at 0° C., was treated with the cyanohydrin 10.06 and heated toreflux for 24 h. The reaction was concentrated in vacuo to yield 10.07,which was used in the next step without purification.

Alternatively 6M HCl prepared by addition of AcCl to dry methanol canalso be used.Step 7

A solution of the amine hydrochloride 10.07 in CH₂Cl₂ (200 mL) wastreated with Et₃N (45.0 mL, 315 mmol) and Boc₂O (45.7 g, 209 mmol) at−78° C. The reaction mixture was then stirred at room temperatureovernight and diluted with HCl (2 M, 200 mL) and extracted into CH₂Cl₂The combined organic layer were dried (MgSO₄) filtered, concentrated invacuo and purified by chromatography (EtOAc/Hex 1:4) to yield hydroxyester 10.08.Step 8.

A solution of methyl ester 10.08 (3 g, 10.5 mmol) in THF/H₂O (1:1) wastreated with LiOH.H₂O (645 mg, 15.75 mmol) and stirred at rt. for 2 h.The reaction mixture was acidified with aq HCl (1 M, 15 mL) andconcentrated in vacuo. The residue was dried in vacuum to afford 10.09in quantitative yield.Step 9

A solution of the acid 10.09 (from above) in CH₂Cl₂ (50 mL) and DMF (25mL) was treated with NH₄Cl (2.94 g, 55.5 mmol), EDCI (3.15 g, 16.5mmol), HOOBt (2.69 g, 16.5 mmol), and NMM (4.4 g, 44 mmol). The reactionmixture was stirred at room temperature for 3 d. The solvents wereremoved under vacuo and the residue was diluted with aq. HCl (250 mL)and extracted with CH₂Cl₂. The combined organic layers were washed withaq. Sat'd. NaHCO₃, dried (MgSO₄) filtered concentrated in vacuo toobtain 10.10, which was used as it was in the following steps.(Alternatively 10.10 can also be obtained directly by the reaction of10.06 (4.5 g, 17.7 mmol) with aq. H₂O₂ (10 mL), LiOH.H₂O (820 mg, 20.8mmol) at 0° C. in 50 mL of CH₃OH for 0.5 h.)Step 10

A solution of 10.10 obtained in the previous step was dissolved in 4 NHCl in dioxane and stirred at rt. for 2 h. The reaction mixture wasconcentrated in vacuo to give the intermediate 10.11 as a solid, whichwas used without further purification.Step 11

The required intermediate 10.12 was obtained from compound 10.09 usingessentially the procedures described above in Steps 9, 10 withappropriate reagents.

Preparation of Intermediate 11.01

Step 1

To a solution of 4-pentyn-1-ol, 11.02 (4.15 g; Aldrich) was addedDess-Martin Periodinane (30.25 g; Aldrich) and the resulting mixture wasstirred for 45 min. before the addition of(tert-Butoxycarbonylmethylene)triphenylphosphorane (26.75 g; Aldrich).The resulting dark reaction was stirred overnight, diluted with EtOAc),washed with aq. sodium sulfite. sat. aq. NaHCO3, water, brine and dried.The volatiles were removed under reduced pressure and the residue waspurified by silica gel column chromatography using 1% EtOAc in hexanesas eluent to give the desired compound, 11.03 (3.92 g). Some impurefractions were also obtained but set aside at this time.Step 2

Using the alkene 11.03 (1.9 g) in n-propanol (20 ml; Aldrich)), benzylcarbamate (4.95 g; Aldrich) in n-propanol (40 ml), NaOH (1.29 g) inwater (79 ml), tert-butyl hypochlorite (3.7 ml), (DHQ)2PHAL (0.423 g;Aldrich)) in n-propanol (37.5 ml), and potassium osmate:dehydrate(0.1544 g; Aldrich) and the procedure set forth in Angew. Chem. Int. Ed.Engl (1998), 35, (23/24), pp. 2813-7. gave a crude product which waspurified by silica gel column chromatography using EtOAc:Hexanes (1:5)to give the desired amino alcohol 11.04 (1.37 g, 37%) as a white solid.Step 3

To the ester 11.04 (0.700 g) was added 4M HCl in dioxane (20 ml;Aldrich) and the resulting mixture was allowed to stand at roomtemperature overnight. The volatiles were removed under reduced pressureto give the acid 11.05 (0.621 g) as a white solid.Step 4

BOP reagent (3.65 g; Sigma) followed by triethylamine (3.45 ml) wereadded to a dichloromethane (20 ml) solution of the carboxylic acid 11.05(2.00 g) and allyl amine (0.616 ml) at room temperature and theresulting mixture was stirred overnight. The reaction mixture waspartitioned between EtOAc and 10% aq. HCl. The organic phase wasseparated, washed with sat. aq. sodium bicarbonate, water, dried(magnesium sulfate). The crude reaction product was purified by silicagel column chromatography using (EtOAc:Hexanes; 70:30) as eluent toprovide the desired amide 11.01 (1.73 g) as a viscous yellow oil.

Preparation of Intermediates 12.03 and 12.04

Step 1

Compound 12.01 was converted to the required material 12.02 usingessentially the procedures described for Intermediate 10.11, Steps 3-8.Step 2

Compound 12.02 was converted to the required intermediate 12.03 usingessentially the procedures described for Intermediate 10.11, Steps 9,10.Step 3

Compound 12.02 was converted to the required intermediate 12.03 usingessentially the procedures described for Intermediate 10.12, Step 11.

Preparation of Intermediate 13.01

Step 1

To a stirred solution of 1-nitrobutane, 13.02 (16.5 g, 0.16 mol) andglyoxylic acid in H₂O (28.1 g, 0.305 mol) and MeOH (122 mL) at 0° C.-5°C., was added dropwise triethylamine (93 mL, 0.667 mol) over 2 hrs. Thesolution was warmed to room temperature, stirred overnight andconcentrated to dryness to give an oil. The oil was then dissolved inH₂O and acidified to pH=1 with 10% HCl, followed by extraction withEtOAc. The combined organic solution was washed with brine, dried overNa₂SO₄, filtered and concentrated to dryness to give the product 13.03(28.1 g, 99% yield).Step 2

To a stirred solution of compound 13.03 (240 g, 1.35 mol) in acetic acid(1.25 L) was added 10% Pd/C (37 g). The resulting solution washydrogenated at 59 psi for 3 hrs and then at 60 psi overnight. Theacetic acid was then evaporated and azeotroped 3 times with toluene,then triturated with MeOH and ether. The solution was then filtered andazeotroped twice with toluene to afford 13.04 as an off white solid (131g, 0.891 mol, 66%).Step 3

To a stirred solution of the amino acid 13.04 (2.0 g, 13.6 mmol) indioxane (10 mL) and H₂O (5 mL) at 0° C., was added 1N NaOH solution (4.3mL, 14.0 mmol). The resulting solution was stirred for 10 minutes,followed by addition of di-t-butyldicarbonate (0.110 g, 14.0 mmol) andstirred at 0° C. for 15 minutes. The solution was then warmed to roomtemperature, stirred for 45 minutes and kept at refrigerator overnightand concentrated to dryness to give a crude material. To the solution ofthis crude material in EtOAc (100 mL) and ice, was added KHSO₄ (3.36 g)and H₂O (32 mL) and stirred for 4-6 minutes. The organic layer was thenseparated and the aqueous layer was extracted twice with EtOAc and thecombined organic layer was washed with water, brine, dried over Na₂SO₄,filtered and concentrated to dryness to give the product 13.05 as aclear gum (3.0 g, 89% yield).Step 4

Compound 13.05 was converted to the required intermediate 13.01 usingessentially the procedures described for Intermediate 10.12, Step 11.

Preparation of Intermediate 14.01

Step 1

Compound 14.02 was converted to the required material 14.03 usingessentially the procedures described for Intermediate 13.01, Steps 1-3.Step 2

Compound 14.03 was converted to the required intermediate 14.01 usingessentially the procedures described for Intermediate 10.12, Step 11.

Preparation of Intermediate 15.01

Step 1

To a suspension of silver nitrite (9 g, 58.5 mmol) in diethyl ether (25mL) at 0° C. was added a solution of 4-iodo-1,1,1-trifluorobutane, 15.02(10 g, 42.0 mmol) in diethyl ether (25 mL) slowly through an additionfunnel (approx. 15 min). The resulting mixture was vigorously stirred at0° C. and warmed to rt. After 50 h, the solid material was filtered offthrough a celite pad. The resulting diethyl ether solution wasconcentrated in vacuo to give 15.03 as colorless oil, which was usedwithout further purification.Step 2

Compound 15.03 was converted to the required material 15.04 usingessentially the procedures described for Intermediate 13.01, Steps 1-3.Step 3

Compound 15.04 was converted to the required intermediate 15.01 usingessentially the procedures described for Intermediate 10.12, Step 11.Preparation of Intermediate 16.01

The acid 16.02 (Winkler, D.; Burger, K., Synthesis, 1996, 1419) isprocessed as described above (preparation of Intermediate 10.12) to givethe expected intermediate 16.01.

Preparation of P₂/P₃—P₂ Moieties

Preparation of Intermediate 20.01

The amino ester 20.01 was prepared following the method of R. Zhang andJ.

S. Madalengoitia (J. Org. Chem. 1999, 64, 330), with the exception thatthe Boc group was cleaved by the reaction of the Boc-protected aminoacid with methanolic HCl (4M HCl in dioxane was also employed for thedeprotection). (Note: In a variation of the reported synthesis, thesulfonium ylide was replaced with the corresponding phosphonium ylide)

Preparation of Intermediate 20.04

Step 1

A solution of commercial amino acid Boc-Chg-OH, 20.02 (Senn chemicals,6.64 g, 24.1 mmol) and amine hydrochloride 20.01 (4.5 g, 22 mmol) inCH₂Cl₂ (100 mL) at 0° C. was treated with BOP reagent and stirred at rt.for 15 h. The reaction mixture was concentrated in vacuo, then it wasdiluted with aq. 1 M HCl and extracted into EtOAc (3×200 mL). Thecombined organic layers were washed with sat'd. NaHCO₃ (200 mL), dried(MgSO₄), filtered and concentrated in vacuo, and chromatographed (SiO₂,EtOAc/Hex 3:7) to obtain 20.03 (6.0 g) as a colorless solid.Step 2

A solution of methyl ester 20.03 (4.0 g, 9.79 mmol) in THF/H₂O (1:1) wastreated with LiOH.H₂O (401 mg, 9.79 mmol) and stirred at rt. for 3 h.The reaction mixture was acidified with aq. HCl and concentrated invacuo to obtain the required intermediate, free acid 20.04.

Preparation of Intermediate 20.08

Step 1

A solution of Boc-tert-Leu 20.05 (Fluka, 5.0 g 21.6 mmol) in dryCH₂Cl₂/DMF (50 mL, 1:1) was cooled to 0° C. and treated with the aminesalt 20.01 (5.3 g, 25.7 mmol), NMM (6.5 g, 64.8 mmol) and BOP reagent(11.6 g, 25.7 mmol). The reaction was stirred at rt. for 24 h, dilutedwith aq. HCl (1 M) and extracted with CH₂Cl₂. The combined organiclayers were washed with HCl (aq, 1 M), sat'd. NaHCO₃, brine, dried(MgSO₄), filtered and concentrated in vacuo and purified bychromatography (SiO2, Acetone/Hexane 1:5) to yield 20.06 as a colorlesssolid.Step 2

A solution of methyl ester 20.06 (4.0 g, 10.46 mmol) was dissolved in 4MHCl in dioxane and stirred at rt. for 3 h. The reaction mixture wasconcentrated in vacuo to obtain the amine hydrochloride salt, 20.07which was used without purification.Step 3

A solution of the amine salt 20.07 (840 mg, 2.64 mmol) in THF (14mL)/acetonitrile (2 mL) was cooled to 0° C. 4-Nitrophenylchloroformate(800 mg, 3.96 mmol) was added followed by pyridine (0.64 mL, 7.92 mmol).The reaction was slowly warmed to room temperature over 3 hrs when TLCindicated reaction completion. Diethyl ether (50 mL) was added and theresulting precipitate was filtered off. The filtrate was washed withsaturated ammonium chloride solution (1×), brine (1×), dried (Na₂SO₄)and concentrated. The residue was purified by flash chromatography using20/80 EtOAc/hexanes which afforded 1.15 g of the required intermediate20.08.

Preparation of Intermediate 21.01

Step 1

To a stirred solution of N-Boc-3,4-dehydroproline 21.02 (5.0 g, 23.5mmol), di-tert-butyl dicarbonate (7.5 g, 34.4 mmol), and4-N,N-dimethylaminopyridine (0.40 g, 3.33 mmol) in acetonitrile (100 mL)at room temperature was added triethylamine (5.0 mL, 35.6 mmol). Theresulting solution was stirred at this temperature for 18 h before itwas concentrated in vacuo. The dark brown residue was purified by flashcolumn chromatography eluting with 10-25% EtOAc/hexane to give theproduct 21.03 as a pale yellow oil (5.29 g, 84%).Step 2

To a stirred solution of the dehydroproline derivative 21.03 (10.1 g,37.4 mmol), benzyltriethylammonium chloride (1.60 g, 7.02 mmol) inchloroform (120 mL) at room temperature was added 50% aqueous sodiumhydroxide (120 g). After vigorously stirred at this temperature for 24h, the dark mixture was diluted with CH₂Cl₂ (200 mL) and diethyl ether(600 mL). After the layers were separated, the aqueous solution wasextracted with CH₂Cl₂/Et₂O (1:2, 3×600 mL). The organic solution wasdried (MgSO₄) and concentrated. The residue was purified by flash columnchromatography using 5-20% EtOAc/hexane to afford 9.34 g (71%) of 21.04as an off-white solid.Step 3

The solution of 21.04 (9.34 g, 26.5 mmol) in CH₂Cl₂ (25 mL) and CF₃CO₂H(50 mL) was stirred at room temperature for 4.5 h before it wasconcentrated in vacuo to give a brown residue, 21.05 which was used inStep 4 without further purification.Step 4

Concentrated hydrochloric acid (4.5 mL) was added to a solution of theresidue 21.05 from Step 3 in methanol (70 mL) and the resulting mixturewas warmed to 65° C. in an oil bath. After 18 h, the mixture wasconcentrated in vacuo to give a brown oil 21.01, which was used furtherwithout purification.

Preparation of Intermediate 22.01

Step 1

Potassium bis(trimethylsilyl)amide (158 ml of a 0.5M solution intoluene; 79 mmol) was added to a stirred suspension ofcyclopropyltriphenylphosphonium bromide (33.12 g; 86.4 mmol) inanhydrous tetrahydrofuran (130 ml) and the resulting orange mixture wasstirred under an atmosphere of nitrogen at room temperature for a periodof 1 h., before the addition of the aldehyde 22.02 (9.68 g; 42.2 mmol)in THF (8 ml). The reaction was then refluxed under an atmosphere ofnitrogen for a period of 2 h. After cooling, methanol, diethyl ether andRochelles salt were added. The organic phase was separated, washed withbrine, dried and concentrated under reduced pressure. The crude reactionproduct was purified by silica gel column chromatography usingEtOAc-hexane (1:99) to EtOAc-hexane (5:95) to provide the alkene 22.03(8.47 g) as a yellow oil.Step 2

A solution of 1M HCl in MeOH/MeOAc was prepared by adding 14.2 ml ofacetyl chloride dropwise into cold methanol and diluting the resultingsolution to 200 ml at room temperature.

The carbamate 22.03 (9.49 g; 37.5 mmol) was dissolved in methanol (12ml) and added to 1M HCl in MeOH/MeOAc (150 ml) while cooled in an icebath. The resulting mixture was maintained at this temperature for 1 h.,then the ice bath was removed and stirring continued overnight at roomtemperature. The volatiles were removed under reduced pressure to yielda yellow oil which was used in the next step without purification.

The yellow oil was dissolved in a mixture of THF (30 ml) and MeOH (20ml) and treated with triethylamine (15 ml; 108 mmol) until the solutionwas pH=9-10. After placing in an ice bath, the mixture was treated withN-Boc-Gly-OSu (11.22 g; 41 mmol). The ice bath was withdrawn and thereaction stirred at room temp. for 1 h. The volatiles were removed underreduced pressure and the residue was purified by silica gel columnchromatography using methanol (1-3%) in dichloromethane providing thedesired amide 22.04 (9.09 g).Step 3

The alcohol 22.04 (9.09 g; 33.6 mmol) was dissolved in acetone (118.5ml) and treated with 2,2-dimethoxypropane (37.4 ml; 304 mmol) andBF₃:Et₂O (0.32 ml; 2.6 mmol) and the resulting mixture was stirred atroom temperature for a period of 5.5 h The reaction solution was treatedwith a few drops of triethylamine and the volatiles were removed underreduced pressure. The residue was purified by silica gel columnchromatography using 5-25% EtOAc in hexanes to provide the N,O-acetal22.05 (8.85 g).Step 4

The carbamate 22.05 (8.81 g; 28.4 mmol) was dissolved in acetonitrile(45 ml) and the solution was cooled to −40° C. under an atmosphere ofnitrogen. Pyridine (6.9 ml; 85.3 mmol) followed by nitrosiumtetrafluoroborate (6.63 g; 56.8 mmol) were added and the resultingreaction mixture maintained below 0° C. until TLC indicated that nostarting material remained (approx. 2.25 h.). Pyrrolidine (20 ml; 240mmol) was added and the cooling bath was withdrawn and stirring wascontinued at room temperature for 1 h. and then the volatiles wereremoved under reduced pressure. The residue was quickly passed through apad of silica gel to provide a yellow oil.

The yellow oil was dissolved in anhydrous benzene (220 ml) and palladiumacetate (0.317 g; 1.41 mmol) was added before heating the resultingmixture to reflux, under an atmosphere of nitrogen for a period of 1.5h. After cooling, the volatiles were removed under reduced pressure andthe dark residue was purified by silica gel column chromatography usingEtOAc-hexane (1:4) to provide the I) the trans-pyrrolidinone 22.06 (1.94g) followed by ii) the cis-pyrrolidinone 22.07 (1.97 g).Step 5

Freshly prepared 1M HCl in MeOAc/MeOH (10 ml; as described above) wasadded to the N,O-acetal 22.06 and stirred at room temperature for 1 h.The solvent was removed under reduced pressure and the residue waspurified by silica gel column chromatography using 0-4% MeOH indichloromethane as eluent to provide the desired alcohol 22.08 (1.42 g),a yellow oil.Step 6

To a solution of the lactam 22.08 (1.29 g; 8.44 mmol) in anhydroustetrahydrofuran (55 ml) was added lithium aluminum hydride (2.40 g; 63.2mmol) and the resulting mixture was refluxed for 8 h. After cooling,water, followed by 15% aq. NaOH were added and the resulting mixture wasfiltered through celite and the solid was washed thoroughly with THF andMeOH. The solvent was removed under reduced pressure and the residueredissolved in dichloromethane, dried and concentrated under reducedpressure to provide the pyrrolidine, used without purification.

Hunigs base (4.5 ml; 25.8 mmol) was added to a mixture ofN-Boc-L-tert-Leu-OH (1.76 g; 7.6 mmol), The crude pyrrolidine and HATU(2.89 g; 7.6 mmol) in anhydrous dichloromethane (50 ml) at −60° C.,under an atmosphere of nitrogen. The resulting reaction was allowed tocome to room temperature slowly, overnight. EtOAc was added and theyellow solution was washed with dil. aq. HCl, sat. aq. sodiumbicarbonate, water, brine. The organic layer was dried and concentratedunder reduced pressure. The residue was purified by silica gel columnchromatography using EtOAc:hexanes (1:3) to give the desired amide 22.09(2.00 g).Step 7

The alcohol 22.09 (2.00 g; 5.67 mmol) was dissolved in acetone (116 ml)and cooled in an ice bath for 10 min. This solution was then added to acooled Jones reagent (14.2 ml; approx 2 mmol/ml) and the resultingmixture was stirred at 5 C for 0.5 h and the cooling bath was removed.The reaction was stirred for a further 2 h. at room temp., before addingto sodium sulfate (28.54 g), celite (15 g) in EtOAc (100 ml).Isopropanol (15 ml) was added after 1 min and then stirred for a further10 min. and filtered. The filtrate was concentrated under reducedpressure, providing a brown oil which was dissolved in EtOAc. Thissolution was washed with water, 3% aq. citric acid, brine, dried andconcentrated to provide the desired carboxylic acid 22.01 (1.64 g) as awhite solid.

Preparation of Intermediate 23.01

Step 1

To the mixture of ester 23.02 (6.0 g) and molecular sieve (5.2 g) inanhydrous methylene chloride (35 mL) was added pyrrolidine (5.7 mL,66.36 mmoL). The resulting brown slurry was stirred at room temperatureunder N₂ for 24 h, filtered and washed with anhydrous CH₃CN. Thecombined filtrate was concentrated to yield the desired product, 23.03.Step 2

To a solution of the product 23.03 from proceeding step in CH₃CN (35 mL)was added anhydrous K₂CO₃, methallyl chloride (2.77 g, 30.5 mmoL), NaI(1.07 g, 6.7 mmoL). The resulting slurry was stirred at ambienttemperature under N₂ for 24 h. 50 mL of ice-cold water was addedfollowed by 2N KHSO₄ solution until pH was 1. EtOAc (100 mL) was addedand the mixture was stirred for 0.75 h. Combined organic layer wascollected and washed with brine, dried over MgSO₄, and evaporated toyield the desired product, 23.04.Step 3

The product 23.04 from the preceding step (2.7 g, 8.16 mmoL) wasdissolved in dioxane (20 mL) and treated with freshly prepared 1N LiOH(9 mL). The reaction mixture was stirred at ambient temperature under N₂for 20 h. The reaction mixture was taken in EtOAc and washed with H₂O.The combined aqueous phase was cooled to 0° C. and acidified to pH 1.65using 1N HCl. The turbid mixture was extracted with EtOAc (2×100 mL).Combined organic layer was washed with brine, dried over MgSO₄, andconcentrated to give the desired acid, 23.05 (3.40 g).Step 4

To a suspension of NaBH(OAc)₃ (3.93 g, 18.5 mmoL) in CH₂Cl₂ (55 mL) wasadded a solution of product 23.05 from preceding step in anhydrousCH₂Cl₂ (20 mL) and acetic acid (2 mL). The slurry was stirred at ambienttemperature for 20 h. Ice cold water (100 mL) was added to the slurryand stirred for ½ hr. Organic layer was separated, filtered, dried andevaporated to yield the desired product, 23.06.Step 5

To a solution of the product 23.06 from preceding step (1.9 g) in MeOH(40 mL) was treated with excess of CH₂N₂/Et₂O solution and stirred forovernight. The reaction mixture was concentrated to dryness to yield acrude residue. The residue was chromatographed on silica gel, elutingwith a gradient of EtOAc/hexane to afford 1.07 g of the pure desiredproduct, 23.07.Step 6

To a solution of product 23.07 from preceding step (1.36 g) in anhydrousCH₂Cl₂ (40 mL) was treated with BF₃. Me₂O (0.7 mL). The reaction mixturewas stirred at ambient temperature for 20 h and quenched with sat.NaHCO₃ (30 mL) ad stirred for ½ hr. Organic layer was separated andcombined organic layer was washed with brine, dried over MgSO₄,concentrated to give crude residue. The residue was chromatographed onsilica gel eluting with a gradient of EtOAc/hexane to afford 0.88 g ofthe desired compound, 23.08.Step 7

To a solution of the product 23.08 (0.92 g) from preceding step in MeOH(30 mL) was added 10% Pd/C (0.16 g) at room temperature and hydrogenatedat ambient temperature under 1 atm. Pressure. The reaction mixture wasstirred for 4 h and concentrated to dryness to yield the desiredcompound, 23.01.

Preparation of P₃ Moieties

Preparation of Intermediate 50.01

Step 1

To a solution of 50.02 (15 g) in MeOH (150 mL) was added conc HCl (3-4mL) and the mixture was refluxed for 16 h. The reaction mixture wascooled to room temperature and concentrated. The residue was taken indiethyl ether (250 mL) and washed with cold saturated sodium bicarbonatesolution, and brine. The organic layer was dried (Na₂SO₄) andconcentrated to afford the methyl ester 50.03 (12.98 g) which wascarried forward without further purification.Step 2

The methyl ester 50.03 from above was dissolved in methylene chloride(100 mL) and cooled to −78° C., under nitrogen atmosphere. DIBAL (1.0 Msolution in methylene chloride, 200 mL) was added dropwise over 2 hperiod. The reaction mixture was warmed to room temperature over 16 h.The reaction mixture was cooled to 0° C. and MeOH (5-8 mL) was addeddropwise. A solution of aqueous 10% sodium potassium tartarate (200 mL)was slowly added with stirring. Diluted with methylene chloride (100 mL)and separated the organic layer (along with some white precipitate). Theorganic layer was washed with 1 N HCl (250 mL), brine (200 mL), dried(Na₂SO₄) and concentrated to provide the alcohol 50.04 (11.00 g) as aclear oil.Step 3

The alcohol 50.04 from above was dissolved in methylene chloride (400mL) and cooled to 0° C. under nitrogen atmosphere. PCC (22.2 g) wasadded in portions and the reaction mixture was slowly warmed to roomtemperature over 16 h. The reaction mixture was diluted with diethylether (500 mL) and filtered through a pad of celite. The filtrate wasconcentrated and the residue was taken in diethyl ether (500 mL). Thiswas passed through a pad of silica gel and the filtrate was concentratedto provide the aldehyde 50.05 which was carried forward without furtherpurification.Step 4

The aldehyde 50.05 from above was converted to the desired material50.01 using essentially the method of Chakraborty et. al (Tetrahedron,1995, 51(33), 9179-90).Preparation of Intermediate 51.01

The required intermediate 51.01 was obtained from the aldehyde 51.02using the literature described procedure (T. K. Chakraborty et al.,Tetrahedron, 1995, 51 (33), 9179-90).

Preparation of Specific Examples

Preparation of Example 1007

Step 1

Commercially available compound 1007a (Aldrich Chemical Co., Milwaukee,Wis., USA) was converted to 1007b according to the literature procedure(M. E. Duggan, J. S. Imagire Synthesis 1989, 131-2) in 90% yield. LC-MS:289 (M+H).Step 2

Deprotection of 1007b using 4M HCl in dioxane at room temperature for 3hrs provided 1007c in quantitative yield. This material was used withoutfurther purification.Step 3

Compound 1007d was obtained from appropriate starting materials/reagentsusing the previously described procedures (See preparation ofIntermediate 20.08).

To a solution of 1007d (200 mg, 0.394 mmol) in dichloromethane (10 mL)at 0° C., under nitrogen atmosphere, was added 1007c (115 mg, 0.512mmol) followed by DIPEA (0.22 mL, 1.182 mmol). The reaction wasmaintained at that temperature for 30 min and stored in the freezer(−20° C.) for 48 hrs. The reaction mixture was quenched with saturatedammonium chloride solution and the product was extracted intodichloromethane (3×). The combine organic layers was washed with brine(1×), dried (Na₂SO₄), filtered and concentrated. The crude residue waspurified by flash chromatography using 30/70 acetone/hexanes whichprovided the required compound 1007e in 69% yield. LC-MS: 557 (M+H).Step 4

Hydrolysis of the methyl ester of 1007e to provide the required acid1007f was carried out as described before (see preparation ofIntermediate 20.04, Step 2) with appropriate modifications.Step 5

Coupling reaction of the acid 1007f (0.125 mmol) with the amine salt10.11 was carried out as described before (see preparation ofIntermediate 20.08, Step 1) with modifications (HATU instead of BOP,DIPEA instead of NMM; the reaction was carried out at 0° C. for 15 minand warmed to 10° C. over 24 hrs) and appropriate amounts of thereagents. The crude material obtained after workup, 1007g was carriedforward without purification. LC-MS: 697.2 (M+H).Step 6

To a cold (0° C.) solution of the material from above, 1007g (0.125mmol) in DMSO/toluene (3 mL each) was added EDCI (240 mg, 1.25 mmol)followed by dichloroacetic acid (0.052 mL, 0.625 mmol). After 15 min,the cold bath was removed and the reaction mixture was warmed to roomtemperature over 16 hr. The reaction mixture was diluted with EtOAc (20mL) and washed with aqueous 1N NaHSO₄ (20 mL). The aqueous layer wasseparated and extracted with EtOAc (20 mL). The combined organic layerswas washed with aqueous 1N NaHSO₄ (20 mL), saturated NaHCO₃ (20 mL),brine (20 mL), dried (Na₂SO₄), filtered and concentrated in vacuo. Thecrude residue was purified by flash column chromatography using 40/60acetone/hexanes to provide the required target compound 1007 (57 mg,0.082 mmol, 66% yield). LC-MS: 695.2 (M+H).Preparation of Example 1044

Step 1

Coupling reaction of the acid 1044a, obtained in a similar manner asdescribed for 1007f (see preparation of Example 1007), with the aminesalt 14.01 was carried out as described before (see preparation ofExample 1007, Step 5). The crude material obtained after workup, 1044bwas carried forward without purification. LC-MS: 725.2 (M+H).Step 2:

To a solution of the material from above, 1044b (0.054 mmol) indichloromethane (5 mL) was added Dess-Martin's periodinane (68 mg, 0.16mmol). The reaction mixture was stirred at room temperature, undernitrogen atmosphere, for 4.5 hrs. The reaction mixture was diluted withdichloromethane (10 mL) and washed with aqueous 10% Na₂S₂O₃ (30 mL),saturated NaHCO₃ (30 mL), brine (30 mL), dried (Na₂SO₄), filtered andconcentrated in vacuo. The crude residue was purified by flash columnchromatography using 35/65 acetone/hexanes to provide the requiredtarget compound 1044 (23 mg, 0.032 mmol, 59% yield). LC-MS: 723.2 (M+H).

Compounds in the following Table 1 and Table 1A were essentiallyprepared using the above-described procedures (Preparation of Examples1007 and 1044) with appropriate reagents and modifications as describedin the General Schemes for Preparation of Target Compounds, Methods A-E.TABLE 1 Example Ki* Mol # Structure (nM) Weight 1001

A 696.885 1002

A 662.868 1003

A 648.841 1004

A 682.859 1005

A 634.815 1006

A 668.832 1007

A 694.87  1008

A 680.843 1009

A 694.87  1010

A 708.896 1011

A 734.934 1012

A 722.923 1013

A 694.87  1014

A 708.896 1015

A 776.895 1016

A 696.885 1017

A 662.868 1018

A 676.895 1019

A 632.799 1020

A 646.826 1021

A 660.852 1022

A 720.907 1023

A 708.896 1024

A 680.843 1025

A 694.87  1026

A 762.868 1027

A 646.826 1028

A 660.852 1029

A 674.879 1030

A 748.961 1031

A 736.95  1032

A 708.896 1033

A 722.923 1034

A 694.87  1035

A 680.843 1036

A 668.832 1037

B 708.896 1038

B 720.907 1039

A 706.881 1040

A 694.87  1041

A 666.816 1042

A 680.843 1043

A 708.896 1044

A 722.923 1045

A 736.95  1046

A 750.977 1047

A 748.841 1048

A 674.879 1049

A 688.906 1050

B 686.89  1051

A 648.841 1052

A 620.788 1053

A 634.815 1054

A 646.826 1055

A 660.852 1056

A 688.906 1057

A 702.933 1058

A 674.879 1059

A 634.815 1060

A 606.761 1061

A 620.788 1062

B 726.912 1063

B 740.938 1064

B 696.885 1065

A 720.907 1066

A 706.881 1067

A 696.885 1068

B 724.939 1069

B 754.965 1070

A 734.934 1071

A 710.912 1072

A 710.912 1073

A 718.892 1074

B 724.939 1075

A 648.841 1076

A 634.815 1077

A 620.788 1078

B 694.87  1079

A 720.907 1080

A 668.832 1081

A 660.852 1082

A 646.826 1083

A 728.851 1084

A 686.89  1085

A 688.906 1086

A 684.874 1087

A 674.871 1088

A 660.844 1089

A 702.924 1090

A 698.892 1091

A 686.882 1092

A 688.898 1093

A 700.908 1094

A 688.898 1095

A 712.919 1096

A 716.951 1097

A 702.924 1098

A 714.935 1099

A 714.935 1100

A 700.908 1101

A 710.903

TABLE 1A Example Ki* LC-MS # Structure (nM) (M + H) 1102

A 729.2 1103

B 715.2 1104

A 725.2 1105

A 727.2 1106

A 731.2 1107

A 717.2 1108

A 727.2 1109

A 729.2 1110

B 772.2 1111

B 757.2 1112

A 767.2 1113

B 769.4 1114

A 715.2 1115

A 724.939 1116

A 729.2 1117

A 727.2 1118

A 711.2 1119

B 701.2 1120

A 741.2 1121

A 741.4Ki* range: A = <75 nM, B = 75-250 nM; C = >250 nMPreparation of Example 1441:

Step 1

To a ice cooled solution of 1441a (4.28 g, 10.08 mmol) in anhydrousether (100 mL) was added LAH (1.53 g, 40.32 mmol) and the reactionmixture was allowed to warm to room temperature overnight. The reactionmixture was cooled to 0° C. and EtOAc (3 mL) was added to it, followedby aqueous KHSO₄ (10 g in 25 mL of H₂O). The gummy residue was extractedwith ether (300 mL) and the organic layer was washed with satd. NaHCO₃,followed by 10% aq. KH₂PO₄, brine, dried over MgSO₄, filtered andconcentrated. The crude residue was purified by flash chromatographyover SiO₂ using ethyl acetate/DCM (1:4) to yield 1441b (2.14 g, 92%).Step 2

To a ice cooled solution of 1441b (743 mg, 3.24 mmol) in anhydrouspyridine (10 mL) was added methyl chloroformate (1 mL, 13 mmol),followed by DMAP (1.6 g, 13 mmol) and the reaction mixture was allowedto warm to room temperature over 2 days. The reaction mixture wasconcentrated and EtOAc (100 mL) was added to it followed by 100 mL ofice-cold (5% KH₂PO₄ containing 0.05 volumes of 1M H₃PO₄). The organiclayer was washed with brine and dried over MgSO₄, filtered andconcentrated. The crude was purified by flash chromatography over SiO₂using ethyl acetate/DCM (1:4) to yield 1441c (931 mg, 100% yield).Step 3

1441c was dissolved in 4M HCl in dioxane (10 mL) and concentrated after30 min. Saturated NaHCO₃ (25 mL) was added to an ice-cold solution ofthe crude hydrochloride salt (194 mg, 1 mmol) in CH₂Cl₂ (25 mL). Thereaction mixture was stirred vigorously for 10 min and COCl₂ (1.85 Msolution in PhMe, 4 mL) was added to it and stirring was continued atroom temperature for 1 h. The organic layer was separated, dried overMgSO₄, filtered and concentrated to half the volume to yield 1441d as a0.05 M solution in CH₂Cl₂.Step 4

To a −20° C. solution 1.17 (10.4 g, 28 mmol; obtained by the hydrolysisof 20.06 using the procedure described for Intermediate 20.04, Step 2)in DCM (300 mL) was added HATU (1.05 equiv, 29.4 mmol, 11.2 g), aminesalt, Intermediate 12.03 (1.0 equiv, 28 mmol, 5.48 g). After 10 min at−20° C., DIPEA (3.6 equiv, 100 mmol, 17.4 mL) was added. Reaction wasstirred at this temp for 16 hr. After 16 hr, the reaction was dilutedwith EtOAc and washed successively with NaHCO₃, citric acid (10% w/w)and brine. Organic layer was dried over MgSO₄, filtered and concentratedin vacuo to yield 14 g of the required intermediate 1441e.Step 5

The hydroxyamide 1441e was oxidized to the required ketoamide 1441f in amanner described for Example 1007, Step 6. LC-MS=507 (M+H)⁺Step 6

Deprotection of the t-Boc functionality of 1441f to give the requiredmaterial 1441g was carried out as described for Example 1007, Step 2.Step 7

To a cooled solution (0° C.) of the amine hydrochloride 1441g (20 mg,0.045 mmol) in CH₂Cl₂ (2.0 mL) was added 1441d (1.35 mL, 0.135 mmol),followed by DIPEA (63 μL, 0.4 mmol). The reaction mixture was stirred atroom temperature for 1.2 h, diluted with ethyl acetate (20 mL), washedwith 3% citric acid, brine, dried over MgSO₄, filtered, concentrated andpurified over SiO₂ using EtOAc/DCM (1:9 to 9:1) to yield 1441 (23 mg).LCMS=620.3 (M+H)⁺.

Compounds in the following table (Table 2) were essentially preparedusing the above described procedures (Preparation of Example 1441) withappropriate reagents and modifications as described in the GeneralSchemes for Preparation of Target Compounds, Methods A-E. TABLE 2Example Ki* Mol # Structure (nM) Weight 1401

A 669.817 1402

A 683.843 1403

B 697.87 1404

A 649.826 1405

A 663.853 1406

B 677.88 1407

A 633.784 1408

A 647.81 1409

B 661.837 1410

A 635.799 1411

A 649.826 1412

A 663.853 1413

A 593.719 1414

A 621.773 1415

A 647.81 1416

A 621.773 1417

A 649.826 1418

A 675.864 1419

A 607.746 1420

A 621.773 1421

A 635.799 1422

C 689.891 1423

B 647.81 1424

A 607.746 1425

A 635.799 1426

A 661.837 1427

A 661.837 1428

A 619.757 1429

A 675.864 1430

A 633.784 1431

B 709.881 1432

A 645.795 1433

A 659.821 1434

B 673.848 1435

B 695.854 1436

A 605.73 1437

A 619.757 1438

B 633.784 1439

B 723.908 1440

B 647.81 1441

A 619.757 1442

A 633.784 1443

B 675.864 1444

A 647.81 1445

A 661.837 1446

B 687.875 1447

A 659.821 1448

A 673.848Ki* range A = <75 nM; B = 75-250 nM, C = >250 nMPreparation of Example 1655

Step 1

To a solution of commercially available compound 1655a (Aldrich ChemicalCo., Milwaukee, Wis., USA, 950 mg, 4.38 mmol) in acetonitrile (40 mL) atroom temperature was added methyl iodide (4.63 mL, 74.42 mmol). Silver(I) oxide (1.62 g, 7.01 mmol) was then added under nitrogen atmosphereand the reaction mixture was refluxed for approximately 16 hrs. (Note:The reaction flask was covered with aluminum foil). At this time, thereaction mixture was cooled to room temperature and filtered through apad of celite. The filter cake was rinsed with ethyl acetate severaltimes. The combined filtrate was concentrated and purified by flashcolumn chromatography using 20/80 to 40/60 ethyl acetate/hexanes toafford 720 mg of the expected product, 1655b.Step 2

Conversion of 1655b to compound 1655c proceeded in quantitative yieldusing previously described procedure (Step 2, Example 1007).Step 3

To a solution of compound 1655c (514 mg, 3.08 mmol) in dichloromethane(20 mL) was added saturated sodium bicarbonate solution (20 mL). Thismixture was stirred vigorously and cooled to 0° C. Phosgene (20 wt % intoluene, 6.5 mL) was added dropwise. The reaction mixture was stirredvigorously for 4.5 hrs while maintaining the temperature at or below 5°C. At this time the reaction mixture was poured into a separatory funneland the organic layer was separated. The organic layer was washed withsaturated ammonium chloride solution (1×), water (1×), dried (Na₂SO₄)and concentrated. The residue, 1655d, was diluted with dichloromethane(10 mL) and used further as a 0.308M solution.Step 4

To a cold (0° C.) solution of 1655e (176 mg, 0.5 mmol; 1655e wasprepared as described for Intermediate 20.08, Steps 1 and 2 usingappropriate starting materials) in dichloromethane (4 mL) was added1655d (0.308 M solution, 4.87 mL, 1.5 mmol) followed by DIPEA (0.276 mL,1.5 mmol). The reaction mixture was maintained at 10° C. for 16 hrs. Thereaction was quenched with saturated ammonium chloride solution and theaqueous layer was extracted with dichloromethane (3×). The combinedorganic layer was washed with brine, dried (Na₂SO₄), filtered andconcentrated in vacuo. The crude residue was purified by flash columnchromatography using 20/80 acetone/hexanes to provide the requiredcompound 1655f (240 mg, 100% yield). LC-MS: 480.1 (M+H).Step 5

Compound 1655f from above was converted to the required target compound1655 using the intermediate 10.11 and procedures described above (Steps4-6, Example 1007). LC-MS of 1655=618.1 (M+H).Preparation of Example 1614

Step 1

To a stirred solution of N-Boc-tert-leucinol 1655a (2.0 g, 9.22 mmol),phenol (1.0 g, 10.6 mmol) and ADDP (3.8 g, 15.1 mmol) in CH₂Cl₂ (80 mL)at rt was bubbled argon gas for 15 min. Triphenylphosphine was thenadded in one portion. The resulting solution was stirred at RT for 18 h.The precipitates were filtered off and washed with diethyl ether (2×30mL). The filtrate was concentrated in vacuo. The residue was purified byflash column chromatography eluting with 2-10% EtOAc/hexane to give thedesired product 1614a (0.33 g, 12%).Step 2

Compound 1614a (0.32 g, 1.13 mmol) was dissolved in a 4 M hydrogenchloride solution in p-dioxane (20 mL) and stirred at RT for 3 h. It wasconcentrated in vacuo to give compound 1614b, which was used withoutfurther purification.Step 3

Compound 1614c was prepared from 1614b according to the proceduresdescribed for Example 1655, Step 3.Step 4

The isocyanate 1614c was converted to the target compound 1614 asdescribed in the General Schemes, Method C using the appropriatereagents and Intermediates.Preparation of Example 1610

Step 1

To a stirred suspension of anhydrous magnesium sulfate in anhydrousCH₂Cl₂ (40 mL) at RT was added concentrated sulfuric acid (0.32 mL, 5.76mmol). The mixture was vigorously stirred for 30 min before a solutionof 1610a (2.0 g, 7.90 mmol) in anhydrous CH₂Cl₂ (15 mL) was added. Themixture was then vigorously stirred at RT for 68 h. Saturated NaHCO₃solution (50 mL) was added cautiously, along with CH₂Cl₂ (100 mL) andwater (50 mL). Two phases were separated, and the aqueous layer wasextracted with CH₂Cl₂ (2×100 mL). The combined organic solution wasdried (MgSO₄), filtered and concentrated in vacuo to give 1610b.Step 2

A suspension of compound 1610b and 10% Pd—C in absolute ethanol wasvigorously stirred under a hydrogen atmosphere for 4 h. The catalyst wasfiltered off through a celite pad. The filtrate was concentrated invacuo to afford 1610c which was used without further purification.Step 3

Compound 1610d was prepared from 1610c according to the proceduresdescribed for Example 1655, Step 3.Step 4

The isocyanate 1610d was converted to the target compound 1610 asdescribed in the General Schemes, Method C using the appropriatereagents and Intermediates.Preparation of Example 1620

Step 1

A suspension of the alcohol 1655a (3.46 g, 12.8 mmol), benzyl bromide(10 mL, 84.2 mmol) and silver (I) oxide (5.0 g, 21.6 mmol) inacetonitrile was stirred vigorously at 76° C. in an oil bath overnight(18 h), The solid material was filtered off and the solution wasconcentrated in vacuo. The product was purified by flash columnchromatography eluting with 540% EtOAc/hexane to give the desiredproduct 1620a (0.78 g, 20%).Step 2

Compound 16120b was prepared from 1620a according to the proceduresdescribed for Example 1614, Step 2.Step 3

Compound 1620c was prepared from 1620b according to the proceduresdescribed for Example 1655, Step 3.Step 4

The isocyanate 1620c was converted to the target compound 1620 asdescribed in the General Schemes, Method C using the appropriatereagents and Intermediates.Preparation of Example 1629

Step 1

To 1441e (600 mg) was added 4M HCl in dioxane (25 mL). The reaction wasstirred at room temperature for 30 min. and concentrated to yield awhite solid, 1629a (490 mg), which was carried forward withoutpurification.Step 2

To a cooled (0° C.) solution of compound 1629a (395 mg) in CH₂Cl₂ (25mL) was added Et₃N (0.57 mL), followed by the isocyanate 1629b(Robinson, Ralph P.; Marfat, Anthony. Eur. Pat. Appl. (1991), EP 436333A2 19910710, 53 pp) in a manner described above (Example 1655, Step 4).The crude hydroxyamide obtained was used without purification.

A solution of the crude hydroxyamide in toluene-DMSO (2.0 mL each) wascooled to 0° C. To the reaction mixture was added EDCI.HCl (410.0 mg),followed by dichloroacetic acetic (0.087 mL), after stirring for 2 h atroom temperature, it was diluted with EtOAc, washed with 1N HCl, satd.NaHCO₃, brine, dried over MgSO4, filtered, concentrated to yield a whitesolid which was purified by chromatography over silica gel usingacetone-hexane (40:60) to afford the title compound 1629 (280.0 mg) as awhite solid: Mass spectrum for C33H49N5O6 (611.77); found FAB(M+H)⁺=612.5Preparation of Example 1628

To a solution of compound 1629 (37.0 mg) in MeOH (2.0 mL) was added Pd—C(10% w/v, 5.0 mg) and the reaction was stirred under hydrogen atmospherefor 1 h, filtered through a pad of celite, concentrated and purified bychromatography over silica gel using acetone-hexane (4:6) to yield therequired compound 1628 (22.0 mg) as a white solid. Mass spectrum forC26H43N5O6 (521.65); found FAB (M+H)⁺=522.6.Preparation of Example 1633

The required title compound 1633 was obtained from the isocyanate 1629band compound 1633a (prepared from 1.17 and 10.11) using proceduresdescribed for Example 1629. Mass spectrum for C34H51 N5O6 (625.80);found FAB (M+H)⁺=626.8.Preparation of Example 1632

To a solution of compound 1633 (10.0 mg) in MeOH (2.0 mL) was added Pd—C( 10% w/v, 2.0 mg) and the reaction was stirred under hydrogenatmosphere for 1 h, filtered through a pad of celite, concentrated andpurified by chromatography over silica gel using acetone-hexane (4:6) toyield the title compound 1632 as a white solid (4.2 mg). Mass spectrumfor C27H45N5O6 (535.68); found FAB (M+H)⁺=536.7.Preparation of Example 1647

Step 1

To a stirred solution of the commercially available compound 1647a(Aldrich Chemical Co., Milwaukee, Wis., USA, 250.0 mg) in MeOH was addedtrimethylsilyl diazomethane (2.0 mL, 2M solution in PhMe). After 20 minthe solvent was removed and the crude was redissolved in CH₂Cl₂ (2.0 mL)and Benzyloxymethyl chloride (1.5 equivalent) was added along with Et₃N(1.5 equivalent). The reaction mixture was stirred overnight, dilutedwith EtOAc, washed successively with 5% Na₂S₂O₃, satd. NaHCO₃, 1N HCl,brine, dried over MgSO4, filtered, concentrated to yield a white solidwhich was purified by chromatography over silica gel using EtOAc-hexane(1:3) to afford the compound 1647b (413 mg) as a white solid: Massspectrum for C20H24O4 (328.40); found FAB (M+H)⁺=329.4.Step 2

To a solution of 0.413 g of compound 1647b in MeOH/H₂O (5.0/0.5 mL) wasadded 0.735 g of KOH. The reaction mixture was refluxed overnight,cooled to room temperature and concentrated. The crude was redissolvedin H₂O (10.0 mL) and acidified with 10% aqueous HCl and extracted withCH₂Cl₂, dried over MgSO₄, filtered and concentrated to afford thecorresponding carboxylic acid 1647c (392 mg). The crude was directlyused in the next step. To a solution of 123.2 mg of acid 1647c intoluene (5.0 mL) was added DPPA (0.09 mL) and Et₃N (0.055 mL). Thereaction mixture was heated at 110° C. for 40 min, cooled and washedwith satd. NaHCO₃, dried over MgSO₄, filtered and concentrated to affordthe isocyanate 1647d. The crude obtained was used without purification.Step 3

The isocyanate 1647d was treated with compound 1629a (90.0 mg) in amanner described in Example 1629 to provide the title compound 1647.Mass spectrum for C40H55N5O7 (717.89); found FAB (M+H)⁺=718.8.Preparation of Example 1648

To a solution of compound 1647 in MeOH was added 6N HCl after 30 min,the MeOH was removed and the crude was redissolved in ethyl acetate andwashed with satd. NaHCO₃. The crude was purified by chromatography oversilica gel using acetone-hexane (40:60) to afford the title compound1648 (25.0 mg) as a white solid: Mass spectrum for C32H47N5O6 (597.75);found FAB (M+H)⁺=598.7.

Compounds in the following table (Table 3) were essentially preparedusing the above described procedures (Preparation of Examples 1610,1614, 1620, 1628, 1629, 1632, 1633, 1647, 1648, 1655) with appropriatereagents and modifications as described in the General Schemes forPreparation of Target Compounds, Methods A-E. TABLE 3 Example Ki* Mol #Structure (nM) Weight 1601

B 577.763 1602

B 563.736 1603

A 637.818 1604

A 547.693 1605

A 665.871 1606

A 651.845 1607

A 561.72 1608

B 679.855 1609

B 691.866 1610

A 619.843 1611

A 633.827 1612

A 647.854 1614

B 625.807 1616

C 663.856 1617

B 573.731 1618

B 639.834 1619

B 653.86 1620

B 639.834 1621

A 665.871 1622

C 725.926 1623

B 691.909 1624

A 601.785 1625

B 635.802 1626

A 575.747 1627

A 591.79 1628

A 521.656 1629

B 611.78 1630

A 623.791 1631

A 637.818 1632

A 535.682 1633

B 625.807 1634

B 743.942 1635

A 623.791 1636

B 673.935 1641

A 757.968 1642

A 637.818 1643

B 771.995 1644

A 651.845 1647

B 717.904 1648

B 597.753 1649

A 645.881 1650

B 631.854 1651

A 609.764 1652

A 623.791 1653

B 653.817 1654

A 603.801 1655

A 617.827 1656

A 589.774 1657

A 603.801 1658

B 611.78 1659

A 603.801 1660

A 589.774 1661

B 671.799 1662

B 629.838 1663

B 651.845 1664

B 663.856 1665

B 705.816 1666

A 611.78 1667

A 665.751 1668

B 653.86 1669

A 625.807 1670

A 589.774 1671

B 617.827 1672

A 603.801 1673

B 617.827 1674

B 603.801 1675

C 687.953 1676

B 683.921 1677

A 645.873 1678

B 659.899 1679

B 647.889 1680

A 695.932 1681

C 699.963 1682

B 671.910 1683

A 657.884 1684

B 659.899 1685

B 655.868 1686

C 659.899 1687

A 617.820 1688

A 631.846Ki* range A = <75 nM; B = 75-250 nM; C = >250 nM

The present invention relates to novel HCV protease inhibitors. Thisutility can be manifested in their ability to inhibit the HCV NS2/NS4aserine protease. A general procedure for such demonstration isillustrated by the following in vitro assay.

Assay for HCV Protease Inhibitory Activity:

Spectrophotometric Assay: Spectrophotometric assay for the HCV serineprotease can be performed on the inventive compounds by following theprocedure described by R. Zhang et al, Analytical Biochemistry, 270(1999) 268-275, the disclosure of which is incorporated herein byreference. The assay based on the proteolysis of chromogenic estersubstrates is suitable for the continuous monitoring of HCV NS3 proteaseactivity. The substrates are derived from the P side of the NS5A-NS5Bjunction sequence (Ac-DTEDVVX(Nva), where X=A or P) whose C-terminalcarboxyl groups are esterified with one of four different chromophoricalcohols (3- or 4-nitrophenol, 7-hydroxy-4-methyl-coumarin, or4-phenylazophenol). Illustrated below are the synthesis,characterization and application of these novel spectrophotometric estersubstrates to high throughput screening and detailed kinetic evaluationof HCV NS3 protease inhibitors.

Materials and Methods:

Materials: Chemical reagents for assay related buffers are obtained fromSigma Chemical Company (St. Louis, Mo.). Reagents for peptide synthesiswere from Aldrich Chemicals, Novabiochem (San Diego, Calif.), AppliedBiosystems (Foster City, Calif.) and Perseptive Biosystems (Framingham,Mass.). Peptides are synthesized manually or on an automated ABI model431A synthesizer (from Applied Biosystems). UV/VIS Spectrometer modelLAMBDA 12 was from Perkin Elmer (Norwalk, Conn.) and 96-well UV plateswere obtained from Corning (Corning, N.Y.). The prewarming block can befrom USA Scientific (Ocala, Fla.) and the 96-well plate vortexer is fromLabline Instruments (Melrose Park, Ill.). A Spectramax Plus microtiterplate reader with monochrometer is obtained from Molecular Devices(Sunnyvale, Calif.).

Enzyme Preparation: Recombinant heterodimeric HCV NS3/NS4A protease(strain 1a) is prepared by using the procedures published previously (D.L. Sali et al, Biochemistry, 37 (1998) 3392-3401). Proteinconcentrations are determined by the Biorad dye method using recombinantHCV protease standards previously quantified by amino acid analysis.Prior to assay initiation, the enzyme storage buffer (50 mM sodiumphosphate pH 8.0, 300 mM NaCl, 10% glycerol, 0.05% lauryl maltoside and10 mM DTT) is exchanged for the assay buffer (25 mM MOPS pH 6.5, 300 mMNaCl, 10% glycerol, 0.05% lauryl maltoside, 5 μM EDTA and 5 μM DTT)utilizing a Biorad Bio-Spin P-6 prepacked column.

Substrate Synthesis and Purification: The synthesis of the substrates isdone as reported by R. Zhang et al, (ibid.) and is initiated byanchoring Fmoc-Nva-OH to 2-chlorotrityl chloride resin using a standardprotocol (K. Barlos et al, Int. J. Pept. Protein Res., 37 (1991),513-520). The peptides are subsequently assembled, using Fmoc chemistry,either manually or on an automatic ABI model 431 peptide synthesizer.The N-acetylated and fully protected peptide fragments are cleaved fromthe resin either by 10% acetic acid (HOAc) and 10% trifluoroethanol(TFE) in dichloromethane (DCM) for 30 min, or by 2% trifluoroacetic acid(TFA) in DCM for 10 min. The combined filtrate and DCM wash isevaporated azeotropically (or repeatedly extracted by aqueous Na₂CO₃solution) to remove the acid used in cleavage. The DCM phase is driedover Na₂SO₄ and evaporated.

The ester substrates are assembled using standard acid-alcohol couplingprocedures (K. Holmber et al, Acta Chem. Scand., B33 (1979) 410-412).Peptide fragments are dissolved in anhydrous pyridine (30-60 mg/ml) towhich 10 molar equivalents of chromophore and a catalytic amount (0.1eq.) of para-toluenesulfonic acid (pTSA) were added.Dicyclohexylcarbodiimide (DCC, 3 eq.) is added to initiate the couplingreactions. Product formation is monitored by HPLC and can be found to becomplete following 12-72 hour reaction at room temperature. Pyridinesolvent is evaporated under vacuum and further removed by azeotropicevaporation with toluene. The peptide ester is deprotected with 95% TFAin DCM for two hours and extracted three times with anhydrous ethylether to remove excess chromophore. The deprotected substrate ispurified by reversed phase HPLC on a C3 or C8 column with a 30% to 60%acetonitrile gradient (using six column volumes). The overall yieldfollowing HPLC purification can be approximately 20-30%. The molecularmass can be confirmed by electrospray ionization mass spectroscopy. Thesubstrates are stored in dry powder form under desiccation.

Spectra of Substrates and Products: Spectra of substrates and thecorresponding chromophore products are obtained in the pH 6.5 assaybuffer. Extinction coefficients are determined at the optimal off-peakwavelength in 1-cm cuvettes (340 nm for 3-Np and HMC, 370 nm for PAP and400 nm for 4-Np) using multiple dilutions. The optimal off-peakwavelength is defined as that wavelength yielding the maximum fractionaldifference in absorbance between substrate and product (productOD−substrate OD)/substrate OD).

Protease Assay: HCV protease assays are performed at 30° C. using a 200μl reaction mix in a 96-well microtiter plate. Assay buffer conditions(25 mM MOPS pH 6.5, 300 mM NaCl, 10% glycerol, 0.05% lauryl maltoside, 5μM EDTA and 5 μM DTT) are optimized for the NS3/NS4A heterodimer (D. L.Sali et al, ibid.)). Typically, 150 μl mixtures of buffer, substrate andinhibitor are placed in wells (final concentration of DMSO≦4% v/v) andallowed to preincubate at 30° C. for approximately 3 minutes. Fifty μlsof prewarmed protease (12 nM, 30° C.) in assay buffer, is then used toinitiate the reaction (final volume 200 μl).The plates are monitoredover the length of the assay (60 minutes) for change in absorbance atthe appropriate wavelength (340 nm for 3-Np and HMC, 370 nm for PAP, and400 nm for 4-Np) using a Spectromax Plus microtiter plate readerequipped with a monochrometer (acceptable results can be obtained withplate readers that utilize cutoff filters). Proteolytic cleavage of theester linkage between the Nva and the chromophore is monitored at theappropriate wavelength against a no enzyme blank as a control fornon-enzymatic hydrolysis. The evaluation of substrate kinetic parametersis performed over a 30-fold substrate concentration range (˜6-200 μM).Initial velocities are determined using linear regression and kineticconstants are obtained by fitting the data to the Michaelis-Mentenequation using non-linear regression analysis (Mac Curve Fit 1.1, K.Raner). Turnover numbers (k_(cat)) are calculated assuming the enzyme isfully active.

Evaluation of Inhibitors and Inactivators: The inhibition constants(K_(i)) for the competitive inhibitors Ac-D-(D-Gla)-L-I-(Cha)-C—OH (27),Ac-DTEDVVA(Nva)-OH and Ac-DTEDVVP(Nva)-OH are determined experimentallyat fixed concentrations of enzyme and substrate by plotting v_(o)/v_(i)vs. inhibitor concentration ([I]_(o)) according to the rearrangedMichaelis-Menten equation for competitive inhibition kinetics:v_(o)/v_(i)=1+[I]_(o)/(K_(i)(1+[S]_(o)/K_(m))), where v_(o) is theuninhibited initial velocity, v_(i) is the initial velocity in thepresence of inhibitor at any given inhibitor concentration ([I]_(o)) and[S]_(o) is the substrate concentration used. The resulting data arefitted using linear regression and the resulting slope,1/(K_(i)(1+[S]_(o)/K_(m)), is used to calculate the K_(i) value. The Ki*values of some of the inventive compounds are shown in Table 6 and Table6A below: TABLE 6 Example Ki* # Structure (nM) 1654

12 1655

24 1631

15 1606

16 1025

6.7 1024

7.3 1086

8 1090

9 1085

11 1023

33 1012

40 1056

30 1029

14 1095

8 1098

15

TABLE 6A Example Ki* # Structure (nM) 1104

13 1108

9 1109

27 1112

20 1114

32 1115

7.4 1116

34 1117

19 1120

30 1121

22

While the present invention has been described with in conjunction withthe specific embodiments set forth above, many alternatives,modifications and other variations thereof will be apparent to those ofordinary skill in the art. All such alternatives, modifications andvariations are intended to fall within the spirit and scope of thepresent invention.

1. A compound, or enantiomers, stereoisomers, rotamers, tautomers, orracemates of said compound, or a pharmaceutically acceptable salt,solvate or ester of said compound, said compound having the generalstructure shown in Formula I:

wherein: R¹ is H, OR⁸, NR⁹R¹⁰, or CHR⁹R¹⁰, wherein R⁸, R⁹ and R¹⁰ can bethe same or different, each being independently selected from the groupconsisting of H, alkyl-, alkenyl-, alkynyl-, aryl-, heteroaryl-,cycloalkyl-, heterocyclyl-, arylalkyl-, and heteroarylalkyl; or R¹⁰ isR¹⁴, wherein R¹⁴ is H, alkyl, aryl, heteroaryl, cycloalkyl, alkyl-aryl,alkyl-heteroaryl, aryl-alkyl, alkenyl, alkynyl or heteroaryl-alkyl; Aand M can be the same or different, each being independently selectedfrom R, OR, NHR, NRR′, SR, SO₂R, and halo; or A and M are connected toeach other such that the moiety:

shown above in Formula I forms either a three, four, six, seven oreight-membered cycloalkyl, a four to eight-membered heterocyclyl, a sixto ten-membered aryl, or a five to ten-membered heteroaryl; E is C(H) orC(R); L is C(H), C(R), CH₂C(R), or C(R)CH₂; R, R′, R², and R³ can be thesame or different, each being independently selected from the groupconsisting of H, alkyl-, alkenyl-, alkynyl-, cycloalkyl-, heteroalkyl-,heterocyclyl-, aryl-, heteroaryl-, (cycloalkyl)alkyl-,(heterocyclyl)alkyl-, aryl-alkyl-, and heteroaryl-alkyl-; or alternatelyR and R′ in NRR′ are connected to each other such that NRR′ forms a fourto eight-membered heterocyclyl; and Y is selected from the followingmoieties:

wherein G is NH; and R¹⁵, R¹⁶, R¹⁷, R¹⁸, and R¹⁹ can be the same ordifferent, each being independently selected from the group consistingof H, alkyl, heteroalkyl, alkenyl, heteroalkenyl, alkynyl,heteroalkynyl, cycloalkyl, heterocyclyl, aryl, arylalkyl, heteroaryl,and heteroarylalkyl, or alternately, (i) either R¹⁵ and R¹⁶ areconnected to each other to form a four to eight-membered cyclicstructure, or R¹⁵ and R¹⁹ are connected to each other to form a four toeight-membered cyclic structure, and (ii) likewise, independently, R¹⁷and R¹⁸ are connected to each other to form a three to eight-memberedcycloalkyl or heterocyclyl; wherein each of said alkyl, aryl,heteroaryl, cycloalkyl or heterocyclyl can be unsubstituted oroptionally independently substituted with one or more moieties selectedfrom the group consisting of: hydroxy, alkoxy, aryloxy, thio, alkylthio,arylthio, amino, amido, alkylamino, arylamino, alkylsulfonyl,arylsulfonyl, sulfonamido, alkylsulfonamido, arylsulfonamido, alkyl,aryl, heteroaryl, keto, carboxy, carbalkoxy, carboxamido,alkoxycarbonylamino, alkoxycarbonyloxy, alkylureido, arylureido, halo,cyano, and nitro.
 2. The compound of claim 1, wherein R¹ is NR⁹R¹⁰, andR⁹ is H, R¹⁰ is H, or R¹⁴ wherein R¹⁴ is H, alkyl, aryl, heteroaryl,cycloalkyl, alkyl-aryl, alkyl-heteroaryl, aryl-alkyl, alkenyl, alkynylor heteroaryl-alkyl.
 3. The compound of claim 2, wherein R¹⁴ is selectedfrom the group consisting of:


4. The compound of claim 1, wherein R² is selected from the groupconsisting of the following moieties:


5. The compound of claim 1, wherein R³ is selected from the groupconsisting of:

wherein R³¹ is OH or O-alkyl; and R³² is H, C(O)CH₃, C(O)OtBu orC(O)N(H)tBu.
 6. The compound of claim 5, wherein R³ is selected from thegroup consisting of the following moieties:


7. The compound of claim 1, wherein Y is selected from the followingmoieties:

wherein G=NH; and R¹⁵, R¹⁶, R¹⁷, R¹⁸, and R¹⁹ can be the same ordifferent, each being independently selected from the group consistingof H, alkyl, heteroalkyl, alkenyl, heteroalkenyl, alkynyl,heteroalkynyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl, oralternately, (i) either R¹⁵ and R¹⁶ are directly connected to form afour to eight-membered cyclic structure, or R¹⁵ and R¹⁹ are directlyconnected to form a four to eight-membered cyclic structure, and (ii)likewise, independently, R¹⁷ and R¹⁵ are directly connected to form athree to eight-membered cycloalkyl or heterocyclyl; wherein each of saidalkyl, aryl, heteroaryl, cycloalkyl or heterocyclyl can be unsubstitutedor optionally independently substituted with one or more moietiesselected from the group consisting of: hydroxy, alkoxy, aryloxy, thio,alkylthio, arylthio, amino, amido, alkylamino, arylamino, alkylsulfonyl,arylsulfonyl, sulfonamido, alkylsulfonamido, arylsulfonamido, keto,carboxy, carbalkoxy, carboxamido, alkoxycarbonylamino,alkoxycarbonyloxy, alkylureido, arylureido, halo, cyano, and nitro. 8.The compound of claim 7, wherein G is NH.
 9. The compound of claim 8,wherein

is selected from the group consisting of:

wherein Y³² is selected from the group consisting of:

R¹⁶ is selected from H, methyl, phenyl, benzyl; and R¹⁵ and R¹⁹ maybethe same or different, each being independently selected from thefollowing:

or alternately, the moiety:

is selected from the following moieties:


10. The compound of claim 9, wherein R¹⁶ is H.
 11. The compound of claim1, wherein the moiety:

is selected from the following structures:


12. The compound of claim 11, wherein the moiety:

is selected from the following structures:


13. The compound of claim 12, wherein the moiety:

is selected from the following structures:


14. The compound of claim 1, wherein R¹⁴ is selected from the groupconsisting of:

R² is selected from the group consisting of the following moieties:

R³is selected from the group consisting of the following moieties:

Y is selected from the group consisting of:

wherein G=NH; and the moiety:

is selected from the group consisting of:

R¹⁶═H; and R¹⁵ and R¹⁹ maybe the same or different, and is selected fromone of the following:

or alternately, the moiety:

is represented by one of the following moieties,

and the moiety:

is:


15. A pharmaceutical composition comprising as an active ingredient atleast one compound of claim
 1. 16. The pharmaceutical composition ofclaim 15 for use in treating an infection of hepatitis C virus (“HCV”).17. The pharmaceutical composition of claim 16 additionally comprisingat least one pharmaceutically acceptable carrier.
 18. The pharmaceuticalcomposition of claim 17, additionally containing at least one antiviralagent.
 19. The pharmaceutical composition of claim 18, stilladditionally containing at least one interferon.
 20. The pharmaceuticalcomposition of claim 19, wherein said at least one antiviral agent isribavirin and said at least one interferon is α-interferon or pegylatedinterferon.
 21. A method of treating an infection of HCV, said methodcomprising administering to a patient in need of such treatment apharmaceutical composition which comprises therapeutically effectiveamounts of at least one compound of claim
 1. 22. The method of claim 21,wherein said administration is oral or subcutaneous.
 23. The use of acompound of claim 1 for the manufacture of a medicament to treat aninfection of HCV.
 24. A method of preparing a pharmaceutical compositionfor treating an infection of HCV, said method comprising bringing intointimate physical contact at least one compound of claim 1 and at leastone pharmaceutically acceptable carrier.
 25. A compound exhibiting HCVprotease inhibitory activity, or enantiomers, stereoisomers, rotamers,tautomers, diastereomers, or racemates of said compound, or apharmaceutically acceptable salt, solvate or ester of said compound,said compound being selected from the compounds of structures listedbelow:


26. A pharmaceutical composition for treating disorders associated withthe HCV, said composition comprising therapeutically effective amount ofone or more compounds in claim 25 and a pharmaceutically acceptablecarrier.
 27. The pharmaceutical composition of claim 26, additionallycontaining at least one antiviral agent.
 28. The pharmaceuticalcomposition of claim 27, additionally containing at least one interferonor PEG-interferon alpha conjugate.
 29. The pharmaceutical composition ofclaim 28, wherein said at least one antiviral agent is ribavirin andsaid at least one interferon is α-interferon or pegylated interferon.30. A method of treatment of a hepatitis C virus associated disorder,comprising administering an effective amount of one or more compounds ofclaim
 25. 31. A method of modulating the activity of hepatitis C virus(HCV) protease, comprising contacting HCV protease with one or morecompounds of claim
 25. 32. A method of treating, preventing, orameliorating one or more symptoms of hepatitis C, comprisingadministering a therapeutically effective amount of one or morecompounds of claim
 25. 33. The method of claim 32, wherein the HCVprotease is the NS3/NS4a protease.
 34. The method of claim 33, whereinthe compound or compounds inhibit HCV NS3/NS4a protease.
 35. A method ofmodulating the processing of hepatitis C virus (HCV) polypeptide,comprising contacting a composition containing the HCV polypeptide underconditions in which said polypeptide is processed with one or morecompounds of claim
 25. 36. A method of treating disorders associatedwith the HCV, said method comprising administering to a patient in needof such treatment, a pharmaceutical composition which comprisestherapeutically effective amounts of at least one compound, orenantiomers, stereoisomers, rotamers, tautomers, diastereomers orracemates of said compound, or a pharmaceutically acceptable salt,solvate or ester of said compound, said compound being selected from thefollowing:


37. A compound of claim 1 in purified form.